A genome is an organism’s complete set of DNA or complete genetic makeup, The entire DNA complement. It describes the identity and the sequence of genes of an organism.
Genomics is the study of entire genomes(structure, function, evolution, mapping, and editing of genomes)
Executing the sequencing and analysis of entire human genome enables more rapid and effective identification of disease associated genes and provide drug companies with pre validated targets.
Proteomics is the systematic high-throughput separation and characterization of proteins within biological systems./ large scale study of protein and their functions.
Proteomics measures protein expression directly, not via gene expression, thus achieving better accuracy. Current work uses 2-dimensional polyacrylamide gel electrophoresis(2D- PAGE) and mass spectrometry.
New separation and characterization technologies, such as protein microarray and high throughput chromatography are being developed.
A genome is an organism’s complete set of DNA or complete genetic makeup, The entire DNA complement. It describes the identity and the sequence of genes of an organism.
Genomics is the study of entire genomes(structure, function, evolution, mapping, and editing of genomes)
Executing the sequencing and analysis of entire human genome enables more rapid and effective identification of disease associated genes and provide drug companies with pre validated targets.
Proteomics is the systematic high-throughput separation and characterization of proteins within biological systems./ large scale study of protein and their functions.
Proteomics measures protein expression directly, not via gene expression, thus achieving better accuracy. Current work uses 2-dimensional polyacrylamide gel electrophoresis(2D- PAGE) and mass spectrometry.
New separation and characterization technologies, such as protein microarray and high throughput chromatography are being developed.
In this slide contains definition, types, causes, inducers and inhibitors, complex drug interactions.
Presented by: SUMASHREE AGGIM (Department of pharmacology).
RIPER, anantapur
This presentation provide knowledge about Gene Expression & its regulation in brief.
i hope it gives some information about gene expression in your academic time.
In this presentation mentioned - Lac Operon and its expressor.
Introduction to Applications of Proteomics Science,
Proteomics- Techniques, Applications of proteomics
Presented by
A. Harsha Vardhan Naidu
Department of Pharmacology
The above presentation consist of the definition of microarray, brief history, general principle of the same, the type of scanner that are used to read or to scan the microarray , type of DNA microarray and finally its various apliccation including the role of DNA microaarray in drug discovery.
genotoxicity describes the property of chemical agents that damages the genetic information within a cell causing mutations, which may lead to cancer. While genotoxicity is often confused with mutagenicity, all mutagens are genotoxic, whereas not all genotoxic substances are mutagenic
Introduction to Genetic Variation in GPCR
G-Protein couple Receptor
Genetic variation in GPCRs
V2 Vasopressin Receptor, Thrombroxane Receptor, P2Y 12ADP Receptor, Chemokine Receptor, Biogenic amine receptors
Presented by
R. REKHA
Department of Pharmacology
The basic aspects of drug discovery starts from target discovery and validation further going to lead identification and optimization. In this particular slide discussion is regarding the target discovery and the tools that have been utilized in this process.
Introduction to Immunotherapeutics
Cell mediated & humoral immunity, Immunosuppressants, Immunostimulants
Presented by
G. Sai Swetha
Department of Pharmacology
In this slide contains principle, types, methods and application of Western Blotting Technique.
Presented by: T.NIRANJAN REDDY (Department of pharmacology).
RIPER, anantapur
Target Validation
Introduction,Drug discovery, Target identification and validation, Target validation and techniques
By
Ms. B. Mary Vishali
Department of Pharmacology
In this slide contains definition, types, causes, inducers and inhibitors, complex drug interactions.
Presented by: SUMASHREE AGGIM (Department of pharmacology).
RIPER, anantapur
This presentation provide knowledge about Gene Expression & its regulation in brief.
i hope it gives some information about gene expression in your academic time.
In this presentation mentioned - Lac Operon and its expressor.
Introduction to Applications of Proteomics Science,
Proteomics- Techniques, Applications of proteomics
Presented by
A. Harsha Vardhan Naidu
Department of Pharmacology
The above presentation consist of the definition of microarray, brief history, general principle of the same, the type of scanner that are used to read or to scan the microarray , type of DNA microarray and finally its various apliccation including the role of DNA microaarray in drug discovery.
genotoxicity describes the property of chemical agents that damages the genetic information within a cell causing mutations, which may lead to cancer. While genotoxicity is often confused with mutagenicity, all mutagens are genotoxic, whereas not all genotoxic substances are mutagenic
Introduction to Genetic Variation in GPCR
G-Protein couple Receptor
Genetic variation in GPCRs
V2 Vasopressin Receptor, Thrombroxane Receptor, P2Y 12ADP Receptor, Chemokine Receptor, Biogenic amine receptors
Presented by
R. REKHA
Department of Pharmacology
The basic aspects of drug discovery starts from target discovery and validation further going to lead identification and optimization. In this particular slide discussion is regarding the target discovery and the tools that have been utilized in this process.
Introduction to Immunotherapeutics
Cell mediated & humoral immunity, Immunosuppressants, Immunostimulants
Presented by
G. Sai Swetha
Department of Pharmacology
In this slide contains principle, types, methods and application of Western Blotting Technique.
Presented by: T.NIRANJAN REDDY (Department of pharmacology).
RIPER, anantapur
Target Validation
Introduction,Drug discovery, Target identification and validation, Target validation and techniques
By
Ms. B. Mary Vishali
Department of Pharmacology
Introduction to Analytical Methods In Invitro Techniques, Analytical Methods, In Vitro Techniques, Spectroscopic Techniques, Chromatography, Electrochemical methods, ELISA, RIA, Bioassay, Electrophoresis
Presented by
K. Thanmaya Divya
Department of Pharmacology
Introduction to Analytical Techniques in Phaese III,
Spectrophotometry, Reflectance photometry, Nephelometry & Turbidimetry, Osmometry, Potentiometry, Flowcytometry, Densitometry, Electrophoresis, LC-MS, ICP-MS
Presented by
B. Kranthi Kumar
Department of Pharmacology
In this slide contains analytical techniques in phase-3 clinical trials.
Presented by: KRANTHI KUMAR BONALA (Department of pharmacology).
RIPER, anantapur
In this slide contains Introduction about XRD and there interpretation.
Presented by: Mohumed omar Mahmoud. (Department of pharmaceutics).
RIPER, anantapur.
In this slide contains contents, steps, different and application of PCR and RT-PCR
Presented by: RAMYA NAGARAJU GARI (Department of pharmacology).
RIPER, anantapur
Introduction to Screening Models Of Anti Cancer Drugs
Need for novel anti cancer drugs, In - vitro methods, In - vivo methods, Advantages and disadvantages
Presented by
T. Niranjan Reddy
Department of Pharmacology
In this slide contains introduction, principle, precautions, solution and assay method for vitamin B series.
Presented by: P. VENKATESH (Department of pharmaceutical analysis),
RIPER, anantapur
Introduction on Dissolution,
Important of dissolution studies,
korsmeyer peppas plot for tablet dissolution,
Presented by
RAMY SALIHEEN
Department of Pharmaceutics
In this slide contains the deep explanation of Methods of Determination for Drug-Excipient Compatibility Studies.
Presented by: G.Aravind Kumar (Department of industrial pharmacy),
RIPER, anantapur.
Fast forward genetic mapping provides candidate genes for resistance to fusar...ICRISAT
Fast forward genetic mapping combined with whole genome sequencing (WGS)and bulked segregantan alysis (BSA)approach was used to identify the candidate genes for Fusarium wilt (FW)and sterility mosaic disease (SMD) resistance in pigeonepa.
3 Jul 2014
In this slide contains introduction, genomic materials of virus and testing method of covid 19 by using RT-PCR.
Presented by: R.Rekha (Department of pharmacology),
RIPER, anantapur.
In this slide contains introduction, methods, supporting media for zone electrophoresis.
Presented by: Mary Vishali. (Department of pharmacology),
RIPER, anantapur.
JOURNAL CLUB PRESENTATION (20L81S0402-PA & QA)
Presented by: K VENKATSAI PRASAD (Department of pharmaceutical analysis and quality assurance).RIPER, anantapur
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...
GENE MAPPING & CLONING
1. Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Gene Mapping & Cloning
A Seminar as a part of curricular requirement for
Master of Pharmacy,
I Year - I semester
Presented by
Mary Vishali
(20L81S0104)
Dept of. Pharmacology
Under the guidance of
Dr. K. Soma Sekhar Reddy M.Pharm, Ph.D.
Associate Professor & HOD Pharamacology
2. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Contents
Introduction
Genome Mapping
Genetic Mapping
Physical Mapping
Uses & Limitations of Gene Mapping
Cloning
Gene Cloning
Reproductive Cloning
Therapeutic Cloning
References
2
3. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Introduction
Gene mapping describes the methods used to identify the locus of a
gene and the distances between genes.
The essence of all genome mapping is to place a collection of
molecular markers onto their respective positions on the genome.
Molecular markers come in all forms. Genes can be viewed as one
special type of genetic markers in the construction of genome maps,
and mapped the same way as any other markers.
3
4. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Genome Mapping
Genetic mapping is based on the use of genetic techniques to
construct maps showing the positions of genes and other sequence
features on a genome.
Genetic techniques include cross-breeding experiments or,
Case of humans, the examination of family histories
(pedigrees).
Physical mapping uses molecular biology techniques to examine
DNA molecules directly in order to construct maps showing the
positions of sequence features, including genes.
4
5. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Genetic Mapping
The first steps of building a genetic map are the development of genetic
markers and a mapping population. Since the closer the two markers are
on the chromosome, the more likely they are to be passed on to the next
generation together, therefore the "co-segregation" patterns of all
markers can be used to reconstruct their order. The genotypes of each
genetic marker are recorded for both parents, and in each individual in
the following generations. The quality of the genetic maps is largely
dependent upon these two factors: the number of genetic markers on the
map and the size of the mapping population.
5
6. RIPER
AUTONOMOUS
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
In gene mapping, any sequence feature that can be
faithfully distinguished from the two parents can be used as a
genetic marker. Genes are represented by "traits" that can be
distinguished between two parents.
Their linkage with other genetic markers are calculated same way
as if they are common markers and the actual gene loci are then
bracketed in a region between the two nearest neighbouring
markers.
The entire process is then repeated by looking at more markers
which target that region to map the gene neighbourhood
6
7. RIPER
AUTONOMOUS
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Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
to a higher resolution until a specific causative locus can be identified.
This process is often referred to as "positional cloning", and it is used
extensively in the study of plant species.
7
8. RIPER
AUTONOMOUS
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Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Physical Mapping
Restriction mapping, which locates the relative positions on a DNA
molecule of the recognition sequences for restriction endonucleases;
Fluorescent in situ hybridization (FISH), in which marker locations
are mapped by hybridizing a probe containing the marker to intact
chromosomes;
Sequence tagged site (STS) mapping, in which the positions of short
sequences are mapped by PCR and/or hybridization analysis of
genome fragments.
8
9. RIPER
AUTONOMOUS
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Physical maps
Physical maps can be generated by aligning the restriction maps of
specific pieces of cloned genomic DNA (for instance, in YAC or
BAC vectors) along the chromosomes.
These maps are extremely useful for the purpose of map-based gene
cloning.
9
10. RIPER
AUTONOMOUS
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Physical Mapping
10
11. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 11
12. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Genetic vs. Physical Distance
Map distances based on recombination frequencies are not a direct
measurement of physical distance along a chromosome.
Recombination “hot spots” overestimate physical length.
Low rates in heterochromatin and centromeres underestimate actual
physical length.
12
13. RIPER
AUTONOMOUS
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NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Genetic vs. Physical Distance
13
14. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Uses of Gene Mapping
Identify genes responsible for diseases.
Heritable diseases
Cancer
Identify genes responsible for traits.
Plants or Animals
Disease resistance
Meat or Milk Production
14
15. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Limitations
A map generated by genetic techniques is rarely sufficient for
directing the sequencing phase of a genome project. This is for two
reasons:
The resolution of a genetic map depends on the number of crossovers
that have been scored .
Genes that are several tens of kb apart may appear at the same
position on the genetic map.
15
16. RIPER
AUTONOMOUS
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Genetic maps have limited accuracy .
Presence of recombination hotspots means that crossovers are
more likely to occur at some points rather than at others.
physical mapping techniques has been developed to address this
problem.
16
17. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
What is Cloning
A clone is a genetically identical copy of an organism, and it may be
naturally occurring or created in the lab. Through the process of
asexual reproduction, organisms such as bacteria (and some plants)
create offspring that are genetically identical to the parent. Modern
genetic technology can also be used to create clones.
17
18. RIPER
AUTONOMOUS
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Types of Cloning
Gene Cloning
Reproductive Cloning
Therapeutic Coning
18
19. RIPER
AUTONOMOUS
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NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Gene Cloning
Making multiple copies of a single gene.
The insertion of fragment of DNA carrying a gene into a cloning vector
and subsequent propagation of recombinant DNA molecule into many
copies is known as gene cloning.
19
20. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 20
21. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Basic Steps of Gene Cloning
Construction of recombinant DNA molecule.
Transport of the recombinant DNA to the host cell.
Multiplication of recombinant DNA molecule.
Division of the host cell.
Numerous cell division resulting in the clone.
21
22. RIPER
AUTONOMOUS
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NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Gene cloning requires specialized tool and
techniques
Vehicles: the central compensation of a gene cloning experiments is the
vehicle which transport the gene into the host cell and is responsible for
its replication. To act as a cloning vehicle a DNA molecule must be
capable of entering a host cell and once inside replicating to produce
multiple copies of itself.
Vector: A DNA molecule capable of replication in a host organism into
which gene is inserted to construct a recombinant DNA molecule.
22
23. RIPER
AUTONOMOUS
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Characteristic of a Vector
It must be able to replicate.
There must be some way to introduce vector DNA into cell.
There must be some means of detecting its presence, preferably
planting test in petri dishes.
23
Cloning vector components
Ori
ampR gene
lacZ gene
Restriction gene
24. RIPER
AUTONOMOUS
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Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Plasmid
Characteristics
They are small and contains 3 – 5 kb of DNA.
They are contain a suitable markers (antibiotic resistant).
They contains suitable restrictions sites that can be used for insertion
DNA fragments for cloning.
24
25. RIPER
AUTONOMOUS
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Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Cloning Vector
A plasmid into which the gene of interest is introduced.
Contains a number of specific gene useful in selection.
25
26. RIPER
AUTONOMOUS
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Cloning Vector Components
Ori
amp R gene
lacZ gene
Restriction gene
Replication origin (ori):
Allow plasmid to replicate in the host cell
26
27. RIPER
AUTONOMOUS
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NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Antibiotic resistance (ampR) gene: allow cells to be resistance to
ampicillin (an antibiotic).
Selection for host cells that have resistance.
This, selection for transformation
β – galactosidase (lacZ) gene: enzyme produced will change a clear
substrate called X-gal into a blue product.
27
28. RIPER
AUTONOMOUS
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Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 28
29. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Step – 1 Forming Recombinant DNA
Where would you insert the DNA of interest so that you can “see” it
in the bacterial cell (Assume cells are grown in X-gal)
Ligate the gene of interest into the vector such that it interrupts the
lacZ gene.
Thus β- Galactosidase is not made.
29
30. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Step – 2 Transformation
Transformation recombinant DNA into bacterial cell.
As bacterial cell multiply, the gene of interest will be replicated
with each other.
Bacterial grown in flakes of liquid medium.
Incubate at optimal growing temperature.
30
31. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Step – 3 Selection
Selection: identify colonies of bacteria containing the recombination
DNA with gene of interest.
Possible bacterial clone products :
A. Bacterial without vector
B. Bacteria with vector without gene
C. Bacterial with vector with the gene of interest
Planting taking a sample of the bacteria and growing them on plates.
Plates have a medium containing: antibiotics
X-gal
31
32. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Selection Mechanism: Antibiotic Resistance
Selection for bacterial clones that contains a vector (select for
proper transformation).
Bacteria are grown on Petri plate containing a specific antibiotics.
Vector confers antibiotics resistance to bacteria because the vector
contains an ampR gene.
Only bacterial cells that properly transformed the vector will live
and grow on the plate.
32
33. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Cloning Application: Bt Plants
Bacillus Thuringiensis a bacterium used a biological pesticide.
Bt gene is cloned into plants so that they will be resistant to pests.
33
34. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Reproductive Cloning
Reproductive cloning involves crating and animal that is genetically
identical to a donor animal through somatic cell nuclear transfer.
In reproductive cloning, the newly created embryo is placed back
into the uterine environment where it can implant and develop.
Ex. The sheep Dolly
34
35. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Therapeutic Cloning
In therapeutic cloning an embryo is created in a similar way to that
of the reproductive cloning but the resulting cloned cells remain in a
dish in the lab, they are not implanted into a female uterus.
35
36. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Cloning Application: Flavr savr Tomatoes
First genetically modified procedure.
Genetically modified tomatoes that suppressed a gene responsible
for fruit ripening.
Process required cloning the gene and transformation a reserve
orientation copy which would have inhibitory effects.
36
37. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
References
T.A Brown, Gene Cloning and DNA Analysis an Introduction, Sixth
Edition.
Anil G. Menon, Charles A. Klanke, and Yan Ru Su, Identification of
Disease Genes By Positional Cloning.
Rita M. Cantor, Analysis of Genetic Linkage.
Huntington’s Disease Genetics Department of neurology, Boston
University School of Medicine, Boston, Massachusetts.
37
38. Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721