Holliday Model of DNA Recombination
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Holliday Model of DNA Recombination which predicts about the crossover and non cross over. Provide base for ds break model.
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Holliday model of crossing over
One of the first plausible models to account for the preceding observations was
formulated by Robin Holliday.
The key features of the Holliday model are the formation of heteroduplex DNA; the
creation of a cross bridge; its migration along the two heteroduplex strands,
termed branch migration; the occurrence of mismatch repair; and the
subsequent resolution, or splicing, of the intermediate structure to yield different
typesof recombinant molecules.
TRANSPOSABLE ELEMENTS
This document discusses transposable elements (TEs), which are segments of DNA that can move within genomes. It covers their discovery by Barbara McClintock in corn in the 1940s. TEs are classified into different types based on their structure and mechanism of movement. The document also examines the mechanisms of transposition, mutagenic effects, regulation, and presence of TEs across bacteria, fungi, and eukaryotes like humans. TEs make up a large fraction of genomes and contribute to genetic variation and disease.
Various model of DNA replication
1. There are four main models of DNA replication: rolling circle replication, theta replication, bidirectional replication of linear DNA, and telomere replication.
2. Rolling circle replication involves nicking circular DNA and using one strand as a template to produce multiple copies of the original circular DNA.
3. Theta replication occurs in prokaryotes and involves unwinding circular DNA at an origin of replication and replicating bi-directionally to form a theta-shaped structure.
4. Bidirectional replication of linear DNA involves unwinding DNA at origins of replication and using leading and lagging strand synthesis to replicate in both directions until the ends of the linear genome are reached.
Site specific recombination
Site-specific recombination involves DNA strand exchange between segments with sequence homology, mediated by site-specific recombinases (SSRs). SSRs recognize and bind to short DNA sequences, cleaving the DNA backbone to exchange helices and rejoin strands. They are classified into tyrosine and serine recombinase families based on mechanism. Tyrosine recombinases like Cre and Flp cleave DNA strands staggered by 6-8bp, linking DNA ends to the recombinase. Serine recombinases simultaneously cleave all four strands staggered by 2bp via phosphoserine bonds. SSRs have applications like tracking cell lineage, ablating genes, and inducing gene expression at specific developmental times.
homologus recombination
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.
Transposable elements in Maize And Drosophila
Transposable elements are mobile DNA sequences found in genomes of all organisms. Barbara McClintock discovered transposable elements called Ac and Ds in maize that cause color patterns in corn kernels. Her discovery showed that genes can move within genomes. Experiments with Drosophila revealed another transposable element called P elements that cause hybrid dysgenesis. Transposable elements can provide genetic variation and flexibility that influences evolution.
Recombination
recombination
types of recombination
models of recombination
biological roles of recombination
Complementation test
This document presents information on complementation tests. It defines complementation tests as a method used to determine if two mutations are in the same gene or different genes. It explains that if the mutations are complementary (in different genes), the offspring will show the parental phenotypes, but if they are not complementary (in the same gene), the offspring will show a new phenotype. Three examples of using complementation test results to determine the number of genes involved are provided. The document concludes by citing a reference for more information on assigning mutations to genes using complementation tests.
Recommended
Holliday model of crossing over
One of the first plausible models to account for the preceding observations was
formulated by Robin Holliday.
The key features of the Holliday model are the formation of heteroduplex DNA; the
creation of a cross bridge; its migration along the two heteroduplex strands,
termed branch migration; the occurrence of mismatch repair; and the
subsequent resolution, or splicing, of the intermediate structure to yield different
typesof recombinant molecules.
TRANSPOSABLE ELEMENTS
This document discusses transposable elements (TEs), which are segments of DNA that can move within genomes. It covers their discovery by Barbara McClintock in corn in the 1940s. TEs are classified into different types based on their structure and mechanism of movement. The document also examines the mechanisms of transposition, mutagenic effects, regulation, and presence of TEs across bacteria, fungi, and eukaryotes like humans. TEs make up a large fraction of genomes and contribute to genetic variation and disease.
Various model of DNA replication
1. There are four main models of DNA replication: rolling circle replication, theta replication, bidirectional replication of linear DNA, and telomere replication.
2. Rolling circle replication involves nicking circular DNA and using one strand as a template to produce multiple copies of the original circular DNA.
3. Theta replication occurs in prokaryotes and involves unwinding circular DNA at an origin of replication and replicating bi-directionally to form a theta-shaped structure.
4. Bidirectional replication of linear DNA involves unwinding DNA at origins of replication and using leading and lagging strand synthesis to replicate in both directions until the ends of the linear genome are reached.
Site specific recombination
Site-specific recombination involves DNA strand exchange between segments with sequence homology, mediated by site-specific recombinases (SSRs). SSRs recognize and bind to short DNA sequences, cleaving the DNA backbone to exchange helices and rejoin strands. They are classified into tyrosine and serine recombinase families based on mechanism. Tyrosine recombinases like Cre and Flp cleave DNA strands staggered by 6-8bp, linking DNA ends to the recombinase. Serine recombinases simultaneously cleave all four strands staggered by 2bp via phosphoserine bonds. SSRs have applications like tracking cell lineage, ablating genes, and inducing gene expression at specific developmental times.
homologus recombination
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.
Transposable elements in Maize And Drosophila
Transposable elements are mobile DNA sequences found in genomes of all organisms. Barbara McClintock discovered transposable elements called Ac and Ds in maize that cause color patterns in corn kernels. Her discovery showed that genes can move within genomes. Experiments with Drosophila revealed another transposable element called P elements that cause hybrid dysgenesis. Transposable elements can provide genetic variation and flexibility that influences evolution.
Recombination
recombination
types of recombination
models of recombination
biological roles of recombination
Complementation test
This document presents information on complementation tests. It defines complementation tests as a method used to determine if two mutations are in the same gene or different genes. It explains that if the mutations are complementary (in different genes), the offspring will show the parental phenotypes, but if they are not complementary (in the same gene), the offspring will show a new phenotype. Three examples of using complementation test results to determine the number of genes involved are provided. The document concludes by citing a reference for more information on assigning mutations to genes using complementation tests.
Mitochondrial genome and its manipulation
Mitochondria contain their own DNA and play an essential role in cellular respiration by generating ATP. While small, the mitochondrial genome encodes components of the electron transport chain. Manipulation of the mitochondrial genome holds promise for crop improvement due to maternal inheritance and absence of position effects. However, transforming the mitochondrial genome remains challenging due to difficulties incorporating foreign DNA and a lack of selectable markers. Successful manipulation could generate cytoplasmic male sterility for hybrid seed production.
Homologous recombination
1. The document discusses models of homologous recombination including the Holliday model and the double-strand break repair model. It describes the key steps and proteins involved in each model.
2. Recombination involves the breakage and rejoining of DNA. In eukaryotes, the MRN/X complex processes DNA breaks. The Rad51 and Rad54 proteins then facilitate strand invasion and D-loop formation during homologous pairing.
3. Homologous recombination proteins from bacteria and eukaryotes catalyze different steps of the process. In E. coli, RecBCD introduces breaks and generates single strands for RecA to perform strand exchange, while RuvAB and Ruv
Dna repair
DNA repair mechanisms identify and correct damage to DNA that occurs due to normal cellular processes and environmental factors. There are two main types of DNA damage: endogenous damage caused by normal cellular processes and exogenous damage caused by external agents like UV radiation and chemicals. The main repair mechanisms are base excision repair, nucleotide excision repair, direct repair via photolyases, and error-prone repair systems like SOS repair. Together, these pathways maintain genome integrity by repairing different types of DNA lesions.
DNA TOPOLOGY
DNA topology studies the geometric properties and spatial relationships of DNA that are unaffected by changes in shape or size. It includes phenomena like supercoiling, knots, and catenanes that involve the linking and twisting of the two DNA strands. DNA topology is characterized by parameters like the linking number, which represents the number of times the two strands are twisted around each other. Enzymes called topoisomerases regulate DNA topology by introducing temporary breaks in the DNA strands to allow strand passage and control supercoiling levels.
Genetic recombination mechanism
Lecture slides on genetic recombination for anyone who is interested in molecular biology or who need to study genetics aspects of biology.
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Second genetic code overlapping and split genes
1) Eukaryotic genes can be organized in complex ways, including overlapping genes where coding sequences partially overlap, and split genes where coding sequences are interrupted by non-coding intron sequences.
2) Overlapping genes were discovered in bacteriophage X174, where the coding sequences of genes D and E overlap but are translated in different reading frames.
3) Split genes have exons, which are the coding sequences included in mRNA, and introns, which are intervening non-coding sequences not included in mRNA. Split genes were first observed in animal viruses in 1977.
Molecular basis of mutations
This document summarizes molecular basis of mutations. It defines mutations as changes in genetic information and describes different types of mutations including point mutations, chromosomal mutations, germline mutations and somatic mutations. It also discusses various mutagens responsible for mutations like chemical mutagens such as alkylating agents, base analogs and reactive oxygen species, and physical mutagens like UV radiation and ionizing radiation. The mechanisms of different mutagens and types of mutations based on their phenotypic effects are also summarized.
Cot curve
Cot value and Cot Curve analysis is a technique for measuring DNA complexity based on renaturation kinetics. DNA is denatured and allowed to reanneal, with larger DNA taking longer. Cot value accounts for DNA concentration, time, and buffer effects, representing repetitive sequences - lower Cot means more repeats. Examples show bacteria have nearly all single-copy DNA, while mouse has varying proportions of single-copy, middle repetitive, and highly repetitive sequences. Cot curve analysis provides information on genome size, complexity, and proportions of sequence types.
Transformation in bacteria
transformation in bacteria is a classical example of horizontal gene transfer which leads to enhanced survivability and also introduction of variations that may lead to evolution
Transposons ppt
Transposons are DNA sequences that can change position within a genome. Barbara McClintock first discovered transposons in corn in the 1940s. There are two classes of transposons: class I (retrotransposons) move via an RNA intermediate, while class II (DNA transposons) move directly via a cut-and-paste mechanism. Transposons make up a large percentage of many genomes and can cause mutations when they insert into genes, which has implications for genetic disease and genome evolution.
Complementation of defined mutations
A complementation test (sometimes called a "cis-trans" test) can be used to test whether the mutations in two strains are in different genes. By taking an example of Benzer's work, complementation has been explained.
Site specific recombination
This document discusses site-specific recombination, including the structures and mechanisms involved. It describes two classes of recombinases - tyrosine recombinases and serine recombinases. Tyrosine recombinases involve cleavage of DNA through formation of a protein-DNA bond using a tyrosine residue. Serine recombinases utilize a phosphoserine bond between DNA and a conserved serine residue. The document provides examples of applications for site-specific recombination such as tracking cell lineage, altering gene expression, and targeted gene knockout.
Transposable elements
Transportable elements are DNA Sequences that move from one location in a chromosome to another within the same chromosome or into another chromosome.
These are DNA Sequences that move from one location in a chromosome to another within the same chromosome or into another chromosome.
These are DNA Sequences that move from one location in a chromosome to another within the same chromosome or into another chromosome.
These are also known as “Jumping genes”.
Tryptophan operon
The tryptophan operon regulates the biosynthesis of tryptophan in E. coli through transcriptional attenuation and repression. It contains five genes encoding the enzymes needed to synthesize tryptophan. When tryptophan levels are high, the tryptophan repressor binds to the operator site, preventing transcription. Additionally, a regulatory region can form a terminator stem-loop structure to halt transcription if tryptophan tRNA levels are high during translation of the leader mRNA sequence. However, if tryptophan levels are low, the terminator structure does not form and transcription of the operon proceeds.
Dna repair mechanisms
DNA repair mechanisms in prokaryotes involve direct repair, excision repair, and mismatch repair. Direct repair converts damaged nucleotides directly back to their original structure using enzymes like photolyase. Excision repair removes damaged sections of DNA through base excision repair which removes single damaged bases using glycosylases and AP endonucleases, or nucleotide excision repair which removes short oligonucleotides. Mismatch repair recognizes and fixes errors made during DNA replication by distinguishing the parental DNA strands and excising the newly synthesized strand containing mistakes.
Restriction Mapping
Restriction enzymes cut DNA molecules at specific recognition sites. Restriction mapping involves digesting an unknown DNA segment with restriction enzymes and analyzing the fragment sizes to determine the locations of restriction sites. One method involves single and double digestions with two enzymes followed by gel electrophoresis to separate the fragments by size. By comparing the fragment patterns between single and double digestions, the positions of each restriction site can be mapped, generating a restriction map of the DNA segment. Restriction mapping was previously important for characterizing cloned DNA but is now easier using DNA sequencing, though analysis of restriction sites remains useful for comparing chromosomal organization between strains.
DNA Replication in eukaryotes and prokaryotes
1. DNA replication is the process by which daughter DNA molecules are synthesized from a parental DNA template. It ensures the genetic information is transferred to the next generation with high fidelity.
2. Replication occurs semi-conservatively such that each new double helix contains one strand from the original parent DNA and one newly synthesized strand. It also occurs bidirectionally from an origin of replication.
3. DNA polymerases are the key enzymes that catalyze DNA synthesis. Other important enzymes and proteins include primase, helicase, topoisomerase, ligase, and single-stranded DNA binding proteins. Together they facilitate the initiation, elongation and termination of DNA replication.
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Homologous recombination
1. The document discusses models of homologous recombination including the Holliday model and the double-strand break repair model. It describes the key steps and proteins involved in each model.
2. Recombination involves the breakage and rejoining of DNA. In eukaryotes, the MRN/X complex processes DNA breaks. The Rad51 and Rad54 proteins then facilitate strand invasion and D-loop formation during homologous pairing.
3. Homologous recombination proteins from bacteria and eukaryotes catalyze different steps of the process. In E. coli, RecBCD introduces breaks and generates single strands for RecA to perform strand exchange, while RuvAB and Ruv
Dna repair
DNA repair mechanisms identify and correct damage to DNA that occurs due to normal cellular processes and environmental factors. There are two main types of DNA damage: endogenous damage caused by normal cellular processes and exogenous damage caused by external agents like UV radiation and chemicals. The main repair mechanisms are base excision repair, nucleotide excision repair, direct repair via photolyases, and error-prone repair systems like SOS repair. Together, these pathways maintain genome integrity by repairing different types of DNA lesions.
DNA TOPOLOGY
DNA topology studies the geometric properties and spatial relationships of DNA that are unaffected by changes in shape or size. It includes phenomena like supercoiling, knots, and catenanes that involve the linking and twisting of the two DNA strands. DNA topology is characterized by parameters like the linking number, which represents the number of times the two strands are twisted around each other. Enzymes called topoisomerases regulate DNA topology by introducing temporary breaks in the DNA strands to allow strand passage and control supercoiling levels.
Genetic recombination mechanism
Lecture slides on genetic recombination for anyone who is interested in molecular biology or who need to study genetics aspects of biology.
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1) Eukaryotic genes can be organized in complex ways, including overlapping genes where coding sequences partially overlap, and split genes where coding sequences are interrupted by non-coding intron sequences.
2) Overlapping genes were discovered in bacteriophage X174, where the coding sequences of genes D and E overlap but are translated in different reading frames.
3) Split genes have exons, which are the coding sequences included in mRNA, and introns, which are intervening non-coding sequences not included in mRNA. Split genes were first observed in animal viruses in 1977.
Molecular basis of mutations
This document summarizes molecular basis of mutations. It defines mutations as changes in genetic information and describes different types of mutations including point mutations, chromosomal mutations, germline mutations and somatic mutations. It also discusses various mutagens responsible for mutations like chemical mutagens such as alkylating agents, base analogs and reactive oxygen species, and physical mutagens like UV radiation and ionizing radiation. The mechanisms of different mutagens and types of mutations based on their phenotypic effects are also summarized.
Cot curve
Cot value and Cot Curve analysis is a technique for measuring DNA complexity based on renaturation kinetics. DNA is denatured and allowed to reanneal, with larger DNA taking longer. Cot value accounts for DNA concentration, time, and buffer effects, representing repetitive sequences - lower Cot means more repeats. Examples show bacteria have nearly all single-copy DNA, while mouse has varying proportions of single-copy, middle repetitive, and highly repetitive sequences. Cot curve analysis provides information on genome size, complexity, and proportions of sequence types.
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transformation in bacteria is a classical example of horizontal gene transfer which leads to enhanced survivability and also introduction of variations that may lead to evolution
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Transposons are DNA sequences that can change position within a genome. Barbara McClintock first discovered transposons in corn in the 1940s. There are two classes of transposons: class I (retrotransposons) move via an RNA intermediate, while class II (DNA transposons) move directly via a cut-and-paste mechanism. Transposons make up a large percentage of many genomes and can cause mutations when they insert into genes, which has implications for genetic disease and genome evolution.
Complementation of defined mutations
A complementation test (sometimes called a "cis-trans" test) can be used to test whether the mutations in two strains are in different genes. By taking an example of Benzer's work, complementation has been explained.
Site specific recombination
This document discusses site-specific recombination, including the structures and mechanisms involved. It describes two classes of recombinases - tyrosine recombinases and serine recombinases. Tyrosine recombinases involve cleavage of DNA through formation of a protein-DNA bond using a tyrosine residue. Serine recombinases utilize a phosphoserine bond between DNA and a conserved serine residue. The document provides examples of applications for site-specific recombination such as tracking cell lineage, altering gene expression, and targeted gene knockout.
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Transportable elements are DNA Sequences that move from one location in a chromosome to another within the same chromosome or into another chromosome.
These are DNA Sequences that move from one location in a chromosome to another within the same chromosome or into another chromosome.
These are DNA Sequences that move from one location in a chromosome to another within the same chromosome or into another chromosome.
These are also known as “Jumping genes”.
Tryptophan operon
The tryptophan operon regulates the biosynthesis of tryptophan in E. coli through transcriptional attenuation and repression. It contains five genes encoding the enzymes needed to synthesize tryptophan. When tryptophan levels are high, the tryptophan repressor binds to the operator site, preventing transcription. Additionally, a regulatory region can form a terminator stem-loop structure to halt transcription if tryptophan tRNA levels are high during translation of the leader mRNA sequence. However, if tryptophan levels are low, the terminator structure does not form and transcription of the operon proceeds.
Dna repair mechanisms
DNA repair mechanisms in prokaryotes involve direct repair, excision repair, and mismatch repair. Direct repair converts damaged nucleotides directly back to their original structure using enzymes like photolyase. Excision repair removes damaged sections of DNA through base excision repair which removes single damaged bases using glycosylases and AP endonucleases, or nucleotide excision repair which removes short oligonucleotides. Mismatch repair recognizes and fixes errors made during DNA replication by distinguishing the parental DNA strands and excising the newly synthesized strand containing mistakes.
Restriction Mapping
Restriction enzymes cut DNA molecules at specific recognition sites. Restriction mapping involves digesting an unknown DNA segment with restriction enzymes and analyzing the fragment sizes to determine the locations of restriction sites. One method involves single and double digestions with two enzymes followed by gel electrophoresis to separate the fragments by size. By comparing the fragment patterns between single and double digestions, the positions of each restriction site can be mapped, generating a restriction map of the DNA segment. Restriction mapping was previously important for characterizing cloned DNA but is now easier using DNA sequencing, though analysis of restriction sites remains useful for comparing chromosomal organization between strains.
DNA Replication in eukaryotes and prokaryotes
1. DNA replication is the process by which daughter DNA molecules are synthesized from a parental DNA template. It ensures the genetic information is transferred to the next generation with high fidelity.
2. Replication occurs semi-conservatively such that each new double helix contains one strand from the original parent DNA and one newly synthesized strand. It also occurs bidirectionally from an origin of replication.
3. DNA polymerases are the key enzymes that catalyze DNA synthesis. Other important enzymes and proteins include primase, helicase, topoisomerase, ligase, and single-stranded DNA binding proteins. Together they facilitate the initiation, elongation and termination of DNA replication.
Tetrad analysis by rk
Tetrad analysis is a technique used to study genetic linkage in fungi and other lower eukaryotes. During meiosis in these organisms, four haploid spores, known as a tetrad, are produced. If spores remain in ordered linear formations, called ordered tetrads, the arrangement allows mapping of genes relative to centromeres. If spores are randomly mixed in unordered tetrads, patterns of allele segregation can determine if two genes are linked. Analysis of tetrad segregation patterns is used to calculate genetic distance between loci.
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Holliday Model of DNA Recombination
- 2. Holliday Model Muhammad Saeed Siddiqui BOT-19-27
- 3. Holiday Model of Recombination • Discovered by Robin Holliday in 1964 in E. Coli • Explain heteroduplex formation and gene conversion during recombination. • Although it has been supplanted by double strand break model, it provides a base for understanding. • It illustrates the critical steps of pairing of homologus duplexes, formation of a heteroduplex, formation of recombination joints, branch migration and Resolution.
- 4. • Duplex • Double stranded DNA molecule, generated by base pairing between complimentary single strands. • Belong to same source • Heteroduplex • Double stranded DNA molecule, generated by base pairing between complimentary single strands. • Double strands derived from different parental duplex molecule. • Occurs during genetic recombination.
- 5. Steps 1. Alignment 2. Nick formation (endo-nucleases) 3. Strand invasion and ligation (heteroduplex formation) 4. Holliday junction/ Holliday fork/ Holliday intermediate/ chi-structure. 5. Branch migration 6. Junction is resolved 7. Recombination
- 6. Alignment
- 7. Nick Formation Nick Endo-Nucleases cuts single strand of each homologus DNA at the same position 5’ 5’ 5’ 5’ 3’ 3’ 3’ 3’
- 8. Chain Invasion • Helicase and SSB Protein, which open the strand and migrate each open strand to the other DNA molecule. Then Ligase enzyme attaches both open strands. 5’ 3’ 3’ 5’ 3’ 5’ 5’ 3’
- 9. Migration • Branch migration occurs which forms the heteroduplex. 5’ 3’ 3’ 5’ 3’ 5’ 5’ 3’
- 10. Resolve of Junction • Duplex pulled away from one another. And rotate at 180 degree. Resolvase (RuvC), Ligase, gyrase and Pol. 5’ 3’ 3’ 5’ 3’ 5’ 3’ 5’
- 11. Animation
- 12. Vertical Cut and Horizontal Cut Non-Crossover Crossover