The S1 nuclease was extracted from Aspergill suoryzae. The S1 nuclease is a specific
single-stranded endonuclease. It can degrade single-stranded DNA and
single-stranded RNA to produce 5'-single-stranded nucleotides or oligonucleotides.
BRIEFLY EXPLAINED PPT ABOUT RESTRICTION ENZYMES, THEIR WORKING SITES, TYPES, ARTIFICIALLY GENERATED RESTRICTION ENZYMES, THEIR MECHANISM OF ACTION, TYPES OF CUTS THEY MAKE, THEIR NOMENCLATURE ETC.
SNP (Single Nucleotide Polymorphic), SNP mapping, SNP profile, SNP types, SNP analysis by gel electropherosis and by mass spectrometry, SNP effects, single strand conformation polymorphism, SNP advantages and disadvantages and application of SNP profile in drug choice
Transcriptomics is the study of RNA, single-stranded nucleic acid, which was not separated from the DNA world until the central dogma was formulated by Francis Crick in 1958, i.e., the idea that genetic information is transcribed from DNA to RNA and then translated from RNA into protein.
In shotgun sequencing the genome is broken randomly into short fragments (1 to 2 kbp long) suitable for sequencing. The fragments are ligated into a suitable vector and then partially sequenced. Around 400–500 bp of sequence can be generated from each fragment in a single sequencing run. In some cases, both ends of a fragment are sequenced. Computerized searching for overlaps between individual sequences then assembles the complete sequence.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Restriction Endonuclease: The Molecular Scissor of DNA - By RIKI NATHRIKI NATH
restriction enducleases are called the molecular scissors of DNA. types of restriction enzymes, their structures, subunits, most importantly the use of Type II restriction endonuclease in recombinant technology, mechanism of enzyme action and their applications.
The S1 nuclease was extracted from Aspergill suoryzae. The S1 nuclease is a specific
single-stranded endonuclease. It can degrade single-stranded DNA and
single-stranded RNA to produce 5'-single-stranded nucleotides or oligonucleotides.
BRIEFLY EXPLAINED PPT ABOUT RESTRICTION ENZYMES, THEIR WORKING SITES, TYPES, ARTIFICIALLY GENERATED RESTRICTION ENZYMES, THEIR MECHANISM OF ACTION, TYPES OF CUTS THEY MAKE, THEIR NOMENCLATURE ETC.
SNP (Single Nucleotide Polymorphic), SNP mapping, SNP profile, SNP types, SNP analysis by gel electropherosis and by mass spectrometry, SNP effects, single strand conformation polymorphism, SNP advantages and disadvantages and application of SNP profile in drug choice
Transcriptomics is the study of RNA, single-stranded nucleic acid, which was not separated from the DNA world until the central dogma was formulated by Francis Crick in 1958, i.e., the idea that genetic information is transcribed from DNA to RNA and then translated from RNA into protein.
In shotgun sequencing the genome is broken randomly into short fragments (1 to 2 kbp long) suitable for sequencing. The fragments are ligated into a suitable vector and then partially sequenced. Around 400–500 bp of sequence can be generated from each fragment in a single sequencing run. In some cases, both ends of a fragment are sequenced. Computerized searching for overlaps between individual sequences then assembles the complete sequence.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Restriction Endonuclease: The Molecular Scissor of DNA - By RIKI NATHRIKI NATH
restriction enducleases are called the molecular scissors of DNA. types of restriction enzymes, their structures, subunits, most importantly the use of Type II restriction endonuclease in recombinant technology, mechanism of enzyme action and their applications.
Biotechnology: Principles and Processes Class XII Chapter 11.pptxBhoomikaDhiman2
Highly descriptive and illustrative presentation based on Biotechnology chapter 11 of NCERT class XII.
This is an important topic especially from biological research point of view.
This is to help students thoroughly understand the topic for exams as well as for future practical applications.
Enzymes that cut DNA at or near specific recognition nucleotide sequences known as restriction sites.
Especial class of enzymes that cleave (cut) DNA at a specific unique internal location along its length.
Often called restriction endonucleases (Because they cut within the molecule).
Discovered in the late 1970s by Werner Arber, Hamilton Smith, and Daniel Nathans.
Essential tools for recombinant DNA technology.
Naturally produced by bacteria that use them as a defense mechanism against viral infection.
Chop up the viral nucleic acids and protect a bacterial cell by hydrolyzing phage DNA.
Assignment on Recombinant DNA Technology and Gene TherapyDeepak Kumar
Assignment on Recombinant DNA Technology and Gene Therapy Basic principles of recombinant DNA technology-Restriction enzymes, various types of vectors, Applications of recombinant DNA technology. Gene therapy- Various types of gene transfer techniques, clinical applications and recent advances in gene therapy
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.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
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.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
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.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
4. ENZYMES
Enzymes are proteins
biological catalysts help drive biochemical reactions.
Enzyme names end with an ase (eg., endonuclease)
Bacteria have evolved a class of enzymes that destroy foreign DNA (eg. Virus DNA).
protect bacteria from bacteriophages (Viruses).
Bacteriophages cannot multiply if their DNA is destroyed by the host.
5. RESTRICTION END/EXO NUCLEASES
Restriction endonucleases RESTRICT viruses
Viral genome is destroyed upon entry.
Restriction endonuclease = Restriction enzymes
Endo (inside), nuclease (cuts nucleic acid)
Exo(outside), nuclease (cuts nucleic acid)
Restriction endonuclease recognizes a short and specific DNA sequence and cuts it from inside.
The specific DNA sequence is called recognition sequence.
6. ORIGINS OF RESTRICTION ENZYMES
1)Bacteria produce restriction enzymes to protect against invading viral DNA/RNA.
9. DISCOVERY
In 1962, Werner Arber, a Swiss biochemist, provided the first evidence for the existence of "molecular scissors" that could cut DNA.
He showed that E. coli bacteria have an enzymatic “immune system” that recognizes and destroys foreign DNA, and modifies native DNA to prevent self- destruction.
10. By the early 1970s these enzymes started to be identified and purified.
It was shown that each species of bacteria had its own population of a SPECIFIC restriction enzyme.
Each enzyme recognized its own specific sequence of DNA bases. It is at this sequence that the DNA was cut.
Smith,Nathans and Arber were awarded the Nobel prise for Physiology and Medicine in 1978 for the discovery of endonucleases.
11. TYPES OF RESTRICTION ENDONULEASES
There are the four distinct types of restriction endonucleases: Type I,Type II, Type III And Type IIs restriction endonucleases.
Type I restriction endonucleases are complex endonucleases and have recognition sequenses of about 15 bp.They cleave the DNA about 1000 bp away from the 5’ end of the sequence “TCA” located within the recognition site , EcoK, EcoB, etc.
12. Type II restriction endonucleases are remarkably stable and induce cleavage either , in most cases within or immediately outside their recognition sequence, which are symmetrical. More then 350 different Type II endonucleases with over 100 different recognition sequences are known. They require Mg+ ions for cleavage.The first Type II enzyme to be isolated was Hind II in 1970.
Only Type II are used for restriction mapping and gene cloning in view of their cleavage only at specific sites.
13. Type III restriction endonucleases are intermediate between the Type I and Type II enzymes.They cleave DNA in the immidiate vicinity of their recognition sites, e.g.,EcoPI, EcoPI5 ,HinfIII, etc.
Type I and Type III restriction enzymes are not used in gene cloning.
The Type IIs enzymes recognize asymmetric target sites, and cleave the DNA duplex on one side of the recognion sequence upto 20 bp away.
14. NOMENCLATURE
Smith and Nathans (1973) proposed enzyme naming scheme
three-letter acronym for each enzyme derived from the source organism
First letter from genus
Next two letters represent species
Additional letter or number represent the strain or serotypes
For example. the enzyme HindII was isolated from Haemophilus influenzae serotype d.
15. FEW RESTRICTION ENZYMES
Enzyme
Organism from which derived
Target sequence (cut at *) 5' -->3'
Bam HI
Bacillus amyloliquefaciens
G* G A T C C
Eco RI
Escherichia coli RY 13
G* A A T T C
Hind III
Haemophilus inflenzae Rd
A* A G C T T
Mbo I
Moraxella bovis
*G A T C
Pst I
Providencia stuartii
C T G C A * G
Sma I
Serratia marcescens
C C C * G G G
Taq I
Thermophilus aquaticus
T * C G A
Xma I
Xanthamonas malvacearum
C * C C G G G
16. R-M SYSTEM
Restriction-modification (R-M) system.
Endonuclease activity: cuts foreign DNA at the recognition site
Methyltransferase activity: protects host DNA from cleavage by the restriction enzyme.
Methyleate one of the bases in each strand
Restriction enzyme and its cognate modification system constitute the R-M system
17. PROTECTION OF SELF DNA
Bacteria protect their self DNA from restriction digestion by methylation of its recognition site.
Methylation is adding a methyl group (CH3) to DNA.
Restriction enzymes are classified based on recognition sequence and methylation pattern.
21. REPELLING BACTERIOPHAGE ATTACK
Take that you wicked virus!
Methylase and restriction endonucleases must recognize the same sequences if they are to function as an effective system
23. Multi-subunit proteins .
Function as a single protein complex .
Contain
two R (restriction) subunits.
two M (methylation) subunits and .
one S (specificity) subunit.
Cleave DNA at random length from recognition site.
24. RECOGNITION SEQUENCES
Each restriction enzyme always cuts at the same recognition sequence.
Produce the same gel banding pattern (fingerprint).
Many restriction sequences are palindromic. For example.
(Read the same in the opposite direction (eg. madam, race car…)
5’ GAATTC 3’
3’ CTTAAG 5’
25. RESTRICTION ENZYME ECORI
Eco RI recognizes the sequence 5’….GAATTC…..
A cut is made between the G and the A on each strand.
This restriction enzyme cleaves the nucleotides 5’AATT overhanging.
These are known as “sticky ends” because hydrogen bonds are available to “stick” to a complimentary 3’TTAA.
Note: Restriction enzymes don’t stop with one cut! They continue to cut at every recognition sequence on a DNA strand.
Restriction Enzyme Cut from EcoRI
26. STICKY END CUTTERS
Most restriction enzymes make staggered cuts.
Staggered cuts produce single stranded “sticky-ends”.
DNA from different sources can be spliced easily because of sticky-end overhangs.
EcoRI
HindIII
27. BLUNT END CUTTERS
Some restriction enzymes cut DNA at opposite base
They leave blunt ended DNA fragments
These are called blunt end cutters
AluI
HaeIII
30. Discovery of enzymes that cut and paste DNA make genetic engineering possible.
Restriction enzyme cuts DNA and generates fragments.
Ligase joins different DNA fragments.
DNA fragments from different species can be ligated (joined) to create Recombinant DNA.
31. DNA FROM TWO SOURCES (RESTRICTION SITES LABELED)
Circular DNA
Linear DNA
37. SOME APPLICATIONS OF RECOMBINANT DNA TECHNOLOGY
Bacteria, Yeasts, and Plants can all be modified to produce important pharmaceuticals, enriched foods, and industrial products.
38. REFERENCES :-
Arber,W. and S.Linn (1969). “ DNA Modification and Restriction”.Annu. Rev. Biochem.,38. pp:467-500.
Reece,J.B., Urry,L.A., Cain,M.L., Wasserman,S.A., Minorsky,P.V. and Robert B. Jackson(2011).“Campbell Biology”, 9th Edition. Pearson Publication, U.S.A.pp:753.
