This document summarizes homologous recombination in eukaryotes and bacteria. In eukaryotes, homologous recombination repairs double-strand DNA breaks through either the double-strand break repair (DSBR) pathway or synthesis-dependent strand annealing (SDSA) pathway. The DSBR pathway forms double Holliday junctions that are resolved to result in crossover or non-crossover products. In bacteria, the RecBCD pathway repairs double-strand breaks and the RecF pathway repairs single-strand gaps. Both pathways involve strand invasion and branch migration to facilitate homologous recombination.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Definition - Rolling circle replication is a process of unidirectional nucleic acid replication.
* can rapidly synthesize multiple copies of circular molecules of DNA or RNA, such as plasmids.
* Eucaryotic also replicate.
* widely used in molecular biology & biomedical
nanotechnology, especially in the field of
biosensing (as a method of signal Amplification).
Steps:
Circular ds DNA will be “nicked”
3` end is elongated →Leading strand
5` end displaced → Lagging strand
made up of double stranded by OKAZAKI fragments.
4) Replication of both “ unnicked” and displaced ss DNA
5) Displaced DNA circulates and synthesis its own complementary strand.
Initation-- phosphate ends, by the action of:
a) Helicase
b) Topoisomerases
c) Single stranded binding proteins(SSBPs)
Elongation-OH group of broken strand, using the unbroken strand as a template. The polymerase will start to move in a circle for elongation, due to which it is named as Rolling Circle Model.
end will be displaced and will grow out like a waving thread.
Termination-* At the point of termination, the linear DNA molecule is cleaved from the circle resulting in a double stranded circular DNA molecule and a single- stranded linear DNA molecule.
* The linear single stranded molecule is circularized by the action of ligase and then replication to double stranded circular plasmid molecule.
Example- Conjugation of F+ and F- bacteria
Diagrammatic representation of Rolling circle
some Examples-Viral DNA
* Human herpes virus
* Human papilloma virus
* Geminivirus
Viral RNA
* pospiviridiae
* Avsunviridiae
Reference:- https://en. m. wikipedia.org
what- when- how.com
https//www.sciencedirect.com
www.slideshare.com
Genetics-notes.wikispace.com
you tube
Prescott 5th edition page.no: 236, 237
Brock biology of microorganism , page.no: 253,616
Mismatch Repair Mechanism Is One Of The Important DNA Repair Mechanism Which Recognizes And Replaces The Wrong Nucleotides. DNA Repair Is Important Since Its Failure Leads To Deadly Diseases Like Cancer. In This Presentation, You Will Learn About DNA Repair, Mismatch Repair, Proteins Involved In Prokaryotic And Eukaryotic MMR, Diagrams, Biological Importance Of MMR And References For Further Study.
Dna supercoiling and role of topoisomerasesYashwanth B S
supercoiling is one of the important process to condenses the huge amount of DNA to fit inside the histone and its also plays a role during the replication ,transcription etc..,these activities is carried out by an enzyme called topoisomerases.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Definition - Rolling circle replication is a process of unidirectional nucleic acid replication.
* can rapidly synthesize multiple copies of circular molecules of DNA or RNA, such as plasmids.
* Eucaryotic also replicate.
* widely used in molecular biology & biomedical
nanotechnology, especially in the field of
biosensing (as a method of signal Amplification).
Steps:
Circular ds DNA will be “nicked”
3` end is elongated →Leading strand
5` end displaced → Lagging strand
made up of double stranded by OKAZAKI fragments.
4) Replication of both “ unnicked” and displaced ss DNA
5) Displaced DNA circulates and synthesis its own complementary strand.
Initation-- phosphate ends, by the action of:
a) Helicase
b) Topoisomerases
c) Single stranded binding proteins(SSBPs)
Elongation-OH group of broken strand, using the unbroken strand as a template. The polymerase will start to move in a circle for elongation, due to which it is named as Rolling Circle Model.
end will be displaced and will grow out like a waving thread.
Termination-* At the point of termination, the linear DNA molecule is cleaved from the circle resulting in a double stranded circular DNA molecule and a single- stranded linear DNA molecule.
* The linear single stranded molecule is circularized by the action of ligase and then replication to double stranded circular plasmid molecule.
Example- Conjugation of F+ and F- bacteria
Diagrammatic representation of Rolling circle
some Examples-Viral DNA
* Human herpes virus
* Human papilloma virus
* Geminivirus
Viral RNA
* pospiviridiae
* Avsunviridiae
Reference:- https://en. m. wikipedia.org
what- when- how.com
https//www.sciencedirect.com
www.slideshare.com
Genetics-notes.wikispace.com
you tube
Prescott 5th edition page.no: 236, 237
Brock biology of microorganism , page.no: 253,616
Mismatch Repair Mechanism Is One Of The Important DNA Repair Mechanism Which Recognizes And Replaces The Wrong Nucleotides. DNA Repair Is Important Since Its Failure Leads To Deadly Diseases Like Cancer. In This Presentation, You Will Learn About DNA Repair, Mismatch Repair, Proteins Involved In Prokaryotic And Eukaryotic MMR, Diagrams, Biological Importance Of MMR And References For Further Study.
Dna supercoiling and role of topoisomerasesYashwanth B S
supercoiling is one of the important process to condenses the huge amount of DNA to fit inside the histone and its also plays a role during the replication ,transcription etc..,these activities is carried out by an enzyme called topoisomerases.
Recombination in repair n damage of DNA.pptxANAKHA JACOB
• Maintaining a low mutation rate is essential for cell viability and health. It is estimated that both in prokaryotic and eukaryotic cells, DNA is replicated with very high fidelity with one wrong nucleotide incorporated once per 108–1010 nucleotides polymerized. The fidelity of DNA replication relies on nucleotide selectivity of replicative DNA polymerase, exonucleolytic proofreading, and post-replicative DNA repair systems.
• Mutations can occur due to errors in DNA replication as well as due to certain damages to the DNA. Errors in replication are corrected to a great extent by proofreading mechanisms. Maintaining the genetic stability that an organism needs for its survival requires not only an extremely accurate mechanism for replicating DNA but also mechanisms for repairing many accidental lesions that occur continually. Most such spontaneous changes in DNA are temporary because they are immediately corrected by a set of processes that are collectively called DNA repair.• Maintaining a low mutation rate is essential for cell viability and health. It is estimated that both in prokaryotic and eukaryotic cells, DNA is replicated with very high fidelity with one wrong nucleotide incorporated once per 108–1010 nucleotides polymerized. The fidelity of DNA replication relies on nucleotide selectivity of replicative DNA polymerase, exonucleolytic proofreading, and post-replicative DNA repair systems.
• Mutations can occur due to errors in DNA replication as well as due to certain damages to the DNA. Errors in replication are corrected to a great extent by proofreading mechanisms. Maintaining the genetic stability that an organism needs for its survival requires not only an extremely accurate mechanism for replicating DNA but also mechanisms for repairing many accidental lesions that occur continually. Most such spontaneous changes in DNA are temporary because they are immediately corrected by a set of processes that are collectively called DNA repair.• Maintaining a low mutation rate is essential for cell viability and health. It is estimated that both in prokaryotic and eukaryotic cells, DNA is replicated with very high fidelity with one wrong nucleotide incorporated once per 108–1010 nucleotides polymerized. The fidelity of DNA replication relies on nucleotide selectivity of replicative DNA polymerase, exonucleolytic proofreading, and post-replicative DNA repair systems.
• Mutations can occur due to errors in DNA replication as well as due to certain damages to the DNA. Errors in replication are corrected to a great extent by proofreading mechanisms. Maintaining the genetic stability that an organism needs for its survival requires not only an extremely accurate mechanism for replicating DNA but also mechanisms for repairing many accidental lesions that occur continually. Most such spontaneous changes in DNA are temporary because they are immediately corrected by a set of processes that are collectively called DNA repair.
Replication:
DNA replication is the biological process of producing two identical copies of DNA from the original/parentral DNA molecule.
This process occurs in all living organism.
Basis for biological inheritance
DNA Replication Is Semiconservative
Replication Begins at an Origin and Usually Proceeds Bidirectionally
DNA Synthesis Proceeds in a 5’-3’ Direction and Is semidiscontinuous
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.
The increased availability of biomedical data, particularly in the public domain, offers the opportunity to better understand human health and to develop effective therapeutics for a wide range of unmet medical needs. However, data scientists remain stymied by the fact that data remain hard to find and to productively reuse because data and their metadata i) are wholly inaccessible, ii) are in non-standard or incompatible representations, iii) do not conform to community standards, and iv) have unclear or highly restricted terms and conditions that preclude legitimate reuse. These limitations require a rethink on data can be made machine and AI-ready - the key motivation behind the FAIR Guiding Principles. Concurrently, while recent efforts have explored the use of deep learning to fuse disparate data into predictive models for a wide range of biomedical applications, these models often fail even when the correct answer is already known, and fail to explain individual predictions in terms that data scientists can appreciate. These limitations suggest that new methods to produce practical artificial intelligence are still needed.
In this talk, I will discuss our work in (1) building an integrative knowledge infrastructure to prepare FAIR and "AI-ready" data and services along with (2) neurosymbolic AI methods to improve the quality of predictions and to generate plausible explanations. Attention is given to standards, platforms, and methods to wrangle knowledge into simple, but effective semantic and latent representations, and to make these available into standards-compliant and discoverable interfaces that can be used in model building, validation, and explanation. Our work, and those of others in the field, creates a baseline for building trustworthy and easy to deploy AI models in biomedicine.
Bio
Dr. Michel Dumontier is the Distinguished Professor of Data Science at Maastricht University, founder and executive director of the Institute of Data Science, and co-founder of the FAIR (Findable, Accessible, Interoperable and Reusable) data principles. His research explores socio-technological approaches for responsible discovery science, which includes collaborative multi-modal knowledge graphs, privacy-preserving distributed data mining, and AI methods for drug discovery and personalized medicine. His work is supported through the Dutch National Research Agenda, the Netherlands Organisation for Scientific Research, Horizon Europe, the European Open Science Cloud, the US National Institutes of Health, and a Marie-Curie Innovative Training Network. He is the editor-in-chief for the journal Data Science and is internationally recognized for his contributions in bioinformatics, biomedical informatics, and semantic technologies including ontologies and linked data.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
2. Introduction
Homologous recombination is a type of genetic
recombination in which nucleotide sequences are exchanged
between two similar or identical molecules of DNA.
It is most widely used by cells to accurately repair harmful
breaks that occur on both strands of DNA, known as double-
strand breaks.
It can also be involved in mutation.
2
4. DSBR Pathway
DSBR pathway is unique because the second 3' overhang
(which was not involved in strand invasion) also forms a
Holliday junction with the homologous chromosome.
The double Holliday junctions are then converted into
recombination products by nicking endonucleases.
Chromosomal crossover is determined by the way of
double Holliday junction is cut, or "resolved".
4
6. SDSA pathway
The invading 3' strand is extended along the recipient DNA duplex
by a DNA polymerase.
Holliday junction between donor and recipient DNA molecules
slides in a process called branch migration.
The newly synthesized 3' end of the invading strand is then able to
anneal to the other 3' overhang in the damaged chromosome through
complementary base pairing.
Ligation process does the work of resealing.
6
8. In Bacteria
Homologous recombination is a major DNA repair process in
bacteria. It is also important for producing genetic diversity in
bacterial populations.
Double-strand DNA breaks in bacteria are repaired by the RecBCD
pathway. Breaks that occur on only one of the two DNA strands,
known as single-strand gaps, are repaired by the RecF pathway.
Both the RecBCD and RecF pathways include a series of reactions
known as branch migration.
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9. RecBCD Pathway
In this pathway, a three-subunit enzyme complex called RecBCD
initiates recombination by binding to a blunt or nearly blunt end of a
break in double-strand DNA. After RecBCD binds the DNA end, the
RecD subunits begin unzipping the DNA duplex through helicase
activity.
The RecB subunit has a nuclease domain, which cuts the single
strand of DNA that emerges from the unzipping process. This
unzipping continues until RecBCD encounters a specific nucleotide
sequence (5'-GCTGGTGG-3') known as a Chi site.
9
10. After reaching chi site enzyme activity decreases to half after a little pause. It is
due to the reason that slower RecB helicase enzyme now in progress in
replacement of faster RecD helicase enzyme.
At chi site RecBCD enzyme cuts the DNA strand with Chi and begins loading
multiple RecA proteins onto the single-stranded DNA.
The resulting RecA-coated nucleoprotein filament then searches out similar
sequences of DNA on a homologous chromosome. Upon finding such a
sequence, the single-stranded nucleoprotein filament moves into the homologous
recipient DNA duplex in a process called strand invasion.
10
12. RecF Pathway
Bacteria appear to use the RecF pathway of homologous recombination to repair
single-strand gaps in DNA. When the RecBCD pathway is inactivated by
mutations, the RecF pathway can also repair DNA double-strand breaks.
In the RecF pathway the RecQ helicase unwinds the DNA and the RecJ
nuclease degrades the strand with a 5’ end, leaving the strand with the 3’ end
intact. RecA protein binds to this strand and is either aided by the RecF, RecO,
and RecR proteins or stabilized by them. The RecA nucleoprotein filament then
searches for a homologous DNA and exchanges places with the identical or
nearly identical strand in the homologous DNA.
12
13. Conclusion
Homologous recombination is conserved across all domains of life, suggesting
that it is a nearly universal biological mechanism.
Homologous recombination that occurs during DNA repair tends to result in
non-crossover products, in effect restoring the damaged DNA molecule as it
existed before the double-strand break.
Homologous recombination is also used in gene targeting, a technique for
introducing genetic changes into target organisms.
13
Editor's Notes
, a type of restriction endonuclease which cuts only one DNA strand.
. Chromosomal crossover will occur if one Holliday junction is cut on the crossing strand and the other Holliday junction is cut on the non-crossing strand.
if the two Holliday junctions are cut on the crossing strands then chromosomes without crossover will be produced.
The invading 3' overhang causes one of the strands of the recipient DNA duplex to be displaced, to form a D-loop. If the D-loop is cut, another swapping of strands forms a cross-shaped structure called a Holliday junction. Resolution of the Holliday junction by some combination of RuvABC or RecG can produce two recombinant DNA molecules with reciprocal genetic types, if the two interacting DNA molecules differ genetically. Alternatively, the invading 3’ end near Chi can prime DNA synthesis and form a replication fork. This type of resolution produces only one type of recombinant (non-reciprocal).