Conjugation and transduction
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Gene Transfer, Types of it, Experiment related to it, Conjugation and its types and how they become F -ve to F +ve and F+ve to Hfr, Transduction and its Types
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transduction
transduction is a process which that bacteriophage is transfer the genetic material to one to another bacterial cell .the transduction is have a two types that is generalized and specialized transduction .the two types of phage will be involve in the transduction process that is virulant and temptate pahge
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
Vectors in biotechnolgy
Vectors are DNA molecules that can carry foreign genes. This document discusses different types of vectors used in biotechnology including plasmid vectors, bacteriophage vectors, cosmid vectors, phagemid vectors, yeast vectors, animal vectors, plant vectors, and artificial chromosomes. It provides details on common vectors like pBR322 and pUC, describing their construction, uses, advantages, and disadvantages.
bacterial transduction
Transduction is the transfer of genetic material between bacteria through bacteriophages. It was discovered in 1952 by Joshua Lederberg and Norton Zinder during their experiments with Salmonella bacteria. There are two types of transduction: generalized transduction, which occurs when a lytic phage picks up bacterial DNA during virus assembly and transfers it to another bacteria; and specialized transduction, which involves the transfer of bacterial DNA adjacent to the phage genome when a temperate phage enters the lysogenic cycle. Transduction contributes to bacterial evolution and genetic diversity by allowing for the lateral transfer of genes between distantly related species.
Conjugation
Conjugation is the transfer of genetic material between bacterial cells that are in direct contact. Joshua Lederberg and Edward Tatum discovered conjugation in 1946 through an experiment where they mixed two auxotrophic E. coli strains that could complement each other's mutations. Bernard Davis provided evidence in 1950 that direct cell contact was required by showing that genetic transfer did not occur when the two strains were separated by a filter. During conjugation, a pilus forms to attach the donor and recipient cells and allows single-stranded DNA to pass between them to make both cells viable donors that can now transfer the genetic material.
Plasmids and types
Autonomously replicating circular fragment present in DNA is called plasmids.
The term plasmid was first introduced by American molecular biologist Joshua Lederberg in1952.
An episome is a plasmid capable of inserting DNA into the host chromosome.
Because of their ability to transfer DNA from one bacterium to another, plasmids are extensively used in recombinant DNA technology or genetic engineering.
Cloning vectors
cloning vectors-
pBR322,
pET21,
pUC 18,
COSMID
SV 40
Lambda phage, M13 Bacteriophage, bacterial artificial chromosome,
yeast artificial chromosome, shuttle vector ,expression vector
Generalized & specialized transduction
This document discusses bacterial transduction, which is a method of gene transfer between bacteria mediated by bacteriophages. There are two main types of transduction: generalized transduction, which occurs during the lytic cycle when viral genes are accidentally incorporated into the new host; and specialized transduction, which involves the specific insertion of viral DNA during lysogeny. Transduction contributes to genetic diversity and evolution in bacteria and can facilitate the spread of antibiotic resistance genes.
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transduction
transduction is a process which that bacteriophage is transfer the genetic material to one to another bacterial cell .the transduction is have a two types that is generalized and specialized transduction .the two types of phage will be involve in the transduction process that is virulant and temptate pahge
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
Vectors in biotechnolgy
Vectors are DNA molecules that can carry foreign genes. This document discusses different types of vectors used in biotechnology including plasmid vectors, bacteriophage vectors, cosmid vectors, phagemid vectors, yeast vectors, animal vectors, plant vectors, and artificial chromosomes. It provides details on common vectors like pBR322 and pUC, describing their construction, uses, advantages, and disadvantages.
bacterial transduction
Transduction is the transfer of genetic material between bacteria through bacteriophages. It was discovered in 1952 by Joshua Lederberg and Norton Zinder during their experiments with Salmonella bacteria. There are two types of transduction: generalized transduction, which occurs when a lytic phage picks up bacterial DNA during virus assembly and transfers it to another bacteria; and specialized transduction, which involves the transfer of bacterial DNA adjacent to the phage genome when a temperate phage enters the lysogenic cycle. Transduction contributes to bacterial evolution and genetic diversity by allowing for the lateral transfer of genes between distantly related species.
Conjugation
Conjugation is the transfer of genetic material between bacterial cells that are in direct contact. Joshua Lederberg and Edward Tatum discovered conjugation in 1946 through an experiment where they mixed two auxotrophic E. coli strains that could complement each other's mutations. Bernard Davis provided evidence in 1950 that direct cell contact was required by showing that genetic transfer did not occur when the two strains were separated by a filter. During conjugation, a pilus forms to attach the donor and recipient cells and allows single-stranded DNA to pass between them to make both cells viable donors that can now transfer the genetic material.
Plasmids and types
Autonomously replicating circular fragment present in DNA is called plasmids.
The term plasmid was first introduced by American molecular biologist Joshua Lederberg in1952.
An episome is a plasmid capable of inserting DNA into the host chromosome.
Because of their ability to transfer DNA from one bacterium to another, plasmids are extensively used in recombinant DNA technology or genetic engineering.
Cloning vectors
cloning vectors-
pBR322,
pET21,
pUC 18,
COSMID
SV 40
Lambda phage, M13 Bacteriophage, bacterial artificial chromosome,
yeast artificial chromosome, shuttle vector ,expression vector
Generalized & specialized transduction
This document discusses bacterial transduction, which is a method of gene transfer between bacteria mediated by bacteriophages. There are two main types of transduction: generalized transduction, which occurs during the lytic cycle when viral genes are accidentally incorporated into the new host; and specialized transduction, which involves the specific insertion of viral DNA during lysogeny. Transduction contributes to genetic diversity and evolution in bacteria and can facilitate the spread of antibiotic resistance genes.
Plasmid
This document discusses plasmids and transposable elements as biotechnology tools. It begins by introducing DNA and plasmids, describing plasmids as extrachromosomal genetic elements found in bacteria. It then classifies plasmids into F, R, and Col factors. Transposable elements are also introduced, including insertion sequences, transposons, and retroelements. Their mechanisms and uses are described, such as mutagenesis and gene tagging. Plasmids are highlighted as important for bacterial conjugation and providing traits like drug resistance. In summary, the document outlines the characteristics and applications of plasmids and transposable elements in biotechnology.
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Rolling circle replication is a process that can rapidly synthesize multiple copies of circular DNA or RNA molecules. It involves the unidirectional replication of circular nucleic acids. The process begins with an initiator protein nicking one strand of the circular DNA. DNA polymerase then uses the 3' end of the nicked strand to initiate replication, displacing the 5' end. Replication continues around the circle to produce a long concatemer of copies. The concatemer is then cleaved and ligated to form multiple double-stranded circular DNA molecules. Rolling circle replication is used by some viruses and plasmids to replicate their genomes and can be harnessed for applications like signal amplification in biosensing.
Bacteriophages
Bacteriophages, or phages, are viruses that infect bacteria. There are two main life cycles for phages: lytic and lysogenic. In the lytic cycle, the phage hijacks the bacterial cell to produce new phages then causes the cell to burst. In the lysogenic cycle, the phage inserts its DNA into the bacterial chromosome where it remains inactive until conditions trigger the lytic cycle. Phages have many applications including phage therapy to treat bacterial infections, using phage lysins as antimicrobials, and phage display to identify molecules that bind to targets of interest.
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1) Griffith discovered a "transforming principle" that allowed non-virulent bacteria to become virulent after exposure to heat-killed virulent bacteria.
2) Avery, MacLeod, and McCarty determined that the transforming principle was DNA through experiments treating components with DNAses, RNAses, and proteases.
3) Hershey and Chase provided definitive evidence that DNA is the genetic material through experiments using bacteriophages containing radioactive DNA or protein to infect bacteria, showing that only DNA was transferred.
P br322
pBR322 is a 4,361 base pair plasmid vector originally constructed in 1977 for use in cloning experiments. It contains genes conferring resistance to ampicillin and tetracycline, which allow selection of recombinant clones, as well as an E. coli origin of replication. Recombinant selection involves insertional inactivation of the tetracycline resistance gene, rendering clones sensitive to tetracycline but resistant to ampicillin. pBR322 was widely used for cloning due to its small size, two selectable markers, and ability to be amplified in host cells. However, it is limited by its mobility between cells and small carrying capacity.
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P uc vectors
pUC vectors are plasmids derived from pBR322 that have a higher copy number of 500-600 copies per cell. They contain an ampicillin resistance gene for selection, as well as the lacZ' gene containing multiple cloning sites. When a gene of interest is inserted, it disrupts the lacZ' gene, allowing for blue-white screening on media containing IPTG and X-gal to identify recombinant colonies that appear white instead of blue. pUC vectors offer advantages over pBR322 such as high copy number and easy selection, though they cannot accommodate inserts larger than 15kb.
Transformation
The document provides information about bacterial transformation. It describes that transformation is the process by which bacteria take up extracellular DNA from their environment. Frederick Griffith first discovered transformation in 1928 while working with pneumococcus bacteria. His experiments showed that a non-virulent rough form could be transformed into a virulent smooth form by DNA from a heat-killed smooth strain. Later experiments by Avery, Macleod and McCarty demonstrated that DNA is the transforming principle and genetic material of bacteria. The document then discusses various methods of bacterial transformation including chemical and physical methods like electroporation and use of calcium chloride. It also explains the molecular mechanism of transformation involving DNA binding, penetration, synapsis formation and integration into the bacterial chromosome.
Prokaryotic genome organization
Prokaryotic genomes are typically organized as single circular chromosomes that are condensed into a nucleoid region within the cell. DNA supercoiling, which involves the over- or under-winding of DNA strands, facilitates compaction of prokaryotic genomes and enables DNA metabolism. Topoisomerases regulate DNA supercoiling by introducing temporary breaks in DNA strands, allowing strands to pass through one another and relieve torsional stress that builds during processes like transcription and replication. The two major types of topoisomerase are types I and II, which introduce single-strand or double-strand breaks, respectively, in regulating supercoiling levels.
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Selection and screening of recombinant clones
This document discusses several methods for selecting recombinant clones after introducing recombinant DNA into host cells:
- Direct selection involves using a gene from the inserted DNA that confers antibiotic resistance to select clones that grow on media containing that antibiotic.
- Insertional inactivation selection works by inactivating a host gene when foreign DNA inserts into it, allowing selection of recombinants.
- Blue-white screening uses a vector with a disrupted lacZ gene; foreign DNA insertion repairs the gene, allowing recombinants to be identified by colony color.
- Colony hybridization detects recombinants by transferring colonies to a membrane and probing for the inserted DNA sequence.
- Immunological tests identify clones expressing antigens encoded by the
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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.
Transposons in bacteria
Barbara McClintock discovered transposons in the 1940s while studying maize. Transposons are segments of DNA that can move within genomes. There are two classes - Class 1 retrotransposons copy and paste RNA into DNA, while Class 2 DNA transposons cut and paste DNA segments. Transposons make up a large portion of many genomes and can cause mutations, but may also create genetic diversity that benefits hosts over time. McClintock received the Nobel Prize for discovering transposons and proposing they play a regulatory role in gene expression.
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This power point presentation is an attempt to present some direct and some indirect evidences in favour of DNA as genetic material. Very few organisms have RNA as genetic material for example plant virus and some bacteriophages
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transduction is a mode of horizontal gene transfer in which the recipient does not come in contact with the donor bacterial cell, it is mediated by temperate phages.
Genetic Recombination
The document discusses various mechanisms of horizontal gene transfer in bacteria, including transformation, transduction, and conjugation. Transformation involves bacteria taking up free DNA from the environment. Transduction involves the transfer of bacterial DNA between cells using bacteriophages. There are two types: generalized transduction transfers random bacterial DNA, while specialized transduction only transfers DNA near the phage insertion site. Conjugation involves the transfer of plasmids between bacteria through direct cell contact via conjugation pili. These mechanisms allow bacteria to rapidly acquire new genes.
genetic variations
Genetic variations can arise in bacteria through several mechanisms:
1. Point mutations or DNA rearrangements can occur spontaneously during DNA replication.
2. DNA can be transferred between bacteria through transformation, transduction, or conjugation. Transformation involves uptake of naked DNA. Transduction involves transfer via bacteriophages. Conjugation involves direct contact between bacteria.
3. These mechanisms allow for acquisition of new genes and increase genetic diversity, which can provide advantages like antibiotic resistance but also influence bacterial pathogenesis.
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Rolling circle replication is a process that can rapidly synthesize multiple copies of circular DNA or RNA molecules. It involves the unidirectional replication of circular nucleic acids. The process begins with an initiator protein nicking one strand of the circular DNA. DNA polymerase then uses the 3' end of the nicked strand to initiate replication, displacing the 5' end. Replication continues around the circle to produce a long concatemer of copies. The concatemer is then cleaved and ligated to form multiple double-stranded circular DNA molecules. Rolling circle replication is used by some viruses and plasmids to replicate their genomes and can be harnessed for applications like signal amplification in biosensing.
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application
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DNA as genetic material
1) Griffith discovered a "transforming principle" that allowed non-virulent bacteria to become virulent after exposure to heat-killed virulent bacteria.
2) Avery, MacLeod, and McCarty determined that the transforming principle was DNA through experiments treating components with DNAses, RNAses, and proteases.
3) Hershey and Chase provided definitive evidence that DNA is the genetic material through experiments using bacteriophages containing radioactive DNA or protein to infect bacteria, showing that only DNA was transferred.
P br322
pBR322 is a 4,361 base pair plasmid vector originally constructed in 1977 for use in cloning experiments. It contains genes conferring resistance to ampicillin and tetracycline, which allow selection of recombinant clones, as well as an E. coli origin of replication. Recombinant selection involves insertional inactivation of the tetracycline resistance gene, rendering clones sensitive to tetracycline but resistant to ampicillin. pBR322 was widely used for cloning due to its small size, two selectable markers, and ability to be amplified in host cells. However, it is limited by its mobility between cells and small carrying capacity.
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P uc vectors
pUC vectors are plasmids derived from pBR322 that have a higher copy number of 500-600 copies per cell. They contain an ampicillin resistance gene for selection, as well as the lacZ' gene containing multiple cloning sites. When a gene of interest is inserted, it disrupts the lacZ' gene, allowing for blue-white screening on media containing IPTG and X-gal to identify recombinant colonies that appear white instead of blue. pUC vectors offer advantages over pBR322 such as high copy number and easy selection, though they cannot accommodate inserts larger than 15kb.
Transformation
The document provides information about bacterial transformation. It describes that transformation is the process by which bacteria take up extracellular DNA from their environment. Frederick Griffith first discovered transformation in 1928 while working with pneumococcus bacteria. His experiments showed that a non-virulent rough form could be transformed into a virulent smooth form by DNA from a heat-killed smooth strain. Later experiments by Avery, Macleod and McCarty demonstrated that DNA is the transforming principle and genetic material of bacteria. The document then discusses various methods of bacterial transformation including chemical and physical methods like electroporation and use of calcium chloride. It also explains the molecular mechanism of transformation involving DNA binding, penetration, synapsis formation and integration into the bacterial chromosome.
Prokaryotic genome organization
Prokaryotic genomes are typically organized as single circular chromosomes that are condensed into a nucleoid region within the cell. DNA supercoiling, which involves the over- or under-winding of DNA strands, facilitates compaction of prokaryotic genomes and enables DNA metabolism. Topoisomerases regulate DNA supercoiling by introducing temporary breaks in DNA strands, allowing strands to pass through one another and relieve torsional stress that builds during processes like transcription and replication. The two major types of topoisomerase are types I and II, which introduce single-strand or double-strand breaks, respectively, in regulating supercoiling levels.
NORTHERN BLOTTING.pptx
This document describes the process of northern blotting. Northern blotting is used to detect specific RNA molecules and analyze gene expression. It involves separating RNA samples by size using gel electrophoresis, transferring them to a membrane, then using a labeled probe to detect the RNA of interest. The key steps are isolating RNA from tissue or cells, separating the RNA by size on a gel, blotting the RNA onto a membrane, hybridizing the membrane with a labeled probe, washing off excess probe, and visualizing where the probe bound to detect the RNA of interest. Northern blotting allows researchers to study gene expression patterns in different tissues, developmental stages, or disease states.
Selection and screening of recombinant clones
This document discusses several methods for selecting recombinant clones after introducing recombinant DNA into host cells:
- Direct selection involves using a gene from the inserted DNA that confers antibiotic resistance to select clones that grow on media containing that antibiotic.
- Insertional inactivation selection works by inactivating a host gene when foreign DNA inserts into it, allowing selection of recombinants.
- Blue-white screening uses a vector with a disrupted lacZ gene; foreign DNA insertion repairs the gene, allowing recombinants to be identified by colony color.
- Colony hybridization detects recombinants by transferring colonies to a membrane and probing for the inserted DNA sequence.
- Immunological tests identify clones expressing antigens encoded by the
DNA Ligation
DNA ligation is the joining of two nucleic acid fragments through the action of an enzyme. Several factors can affect the ligation reaction, including the concentration of enzyme, DNA, and cofactors like ATP or NAD+. The DNA concentration is particularly important, as higher concentrations favor intermolecular ligation between separate DNA molecules, while lower concentrations favor intramolecular ligation where a DNA molecule joins its own ends. DNA ligase carries out ligation through a three-step catalytic mechanism involving adenylation of the enzyme and two phosphoryl transfers.
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.
Transposons in bacteria
Barbara McClintock discovered transposons in the 1940s while studying maize. Transposons are segments of DNA that can move within genomes. There are two classes - Class 1 retrotransposons copy and paste RNA into DNA, while Class 2 DNA transposons cut and paste DNA segments. Transposons make up a large portion of many genomes and can cause mutations, but may also create genetic diversity that benefits hosts over time. McClintock received the Nobel Prize for discovering transposons and proposing they play a regulatory role in gene expression.
Bidirectional and rolling circular dna replication
The document discusses bidirectional and rolling circular DNA replication. It begins by explaining that DNA replication is semi-conservative, with each new DNA molecule composed of one original strand and its complement. It then describes how in bidirectional replication, replication forks move in opposite directions from a single origin to copy both strands simultaneously. Rolling circular replication involves the continuous, unidirectional synthesis of multiple copies of a circular DNA or RNA genome through the sequential steps of initiation by introducing a nick, elongation by polymerase moving in a circle, and termination by cleaving the replicated strand.
Dna as genetic material
This power point presentation is an attempt to present some direct and some indirect evidences in favour of DNA as genetic material. Very few organisms have RNA as genetic material for example plant virus and some bacteriophages
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The document discusses various mechanisms of horizontal gene transfer in bacteria, including transformation, transduction, and conjugation. Transformation involves bacteria taking up free DNA from the environment. Transduction involves the transfer of bacterial DNA between cells using bacteriophages. There are two types: generalized transduction transfers random bacterial DNA, while specialized transduction only transfers DNA near the phage insertion site. Conjugation involves the transfer of plasmids between bacteria through direct cell contact via conjugation pili. These mechanisms allow bacteria to rapidly acquire new genes.
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Genetic variations can arise in bacteria through several mechanisms:
1. Point mutations or DNA rearrangements can occur spontaneously during DNA replication.
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This document summarizes bacterial genetic systems including transformation, conjugation, and transduction. It describes that bacterial genomes are circular and contain chromosomal DNA and plasmids. Plasmids can be conjugative or non-conjugative. Transformation involves the uptake of naked DNA by bacterial cells. Conjugation involves the transfer of genetic material between bacterial cells through direct cell contact via pili. Transduction involves the transfer of bacterial DNA by bacteriophages. These three processes allow for genetic recombination and diversity in bacteria.
ppt of Bacterial genetic system
This document summarizes three mechanisms of genetic transfer in bacteria: transformation, conjugation, and transduction. Transformation involves the uptake of naked DNA by a bacterial cell. Conjugation is the transfer of genetic material between bacteria via cell-to-cell contact through a pilus. Transduction is the process by which bacteriophage viruses transmit DNA between bacteria. These three mechanisms allow for genetic recombination and diversity in bacterial populations.
Genetic transformation in Prokaryotes
Genetic transformation in prokaryotes has led to the discovery of the three major methods of transformationin bacteria i.e transformation, conjugation and transuction whichcommonly uses the bacterial- phages as vectors to transfer dna.
Bacterial resistance mechanisms and new trends for resistance overcoming
This document discusses bacterial resistance and its mechanisms. It begins with an overview of the increasing issue of antimicrobial resistance worldwide. It then covers the origins of resistance as either intrinsic or acquired through mutation or horizontal gene transfer. The major mechanisms of acquired resistance are expressed genes coding for altered drug targets, enzymatic drug inactivation, efflux pumps, and biofilms. Factors promoting resistance include antibiotic misuse in medicine and agriculture as well as a lack of new drug development. The consequences are serious infections that are difficult to treat.
How does horizontal transfer affect the evolution of anti-biotic res.pdf
How does horizontal transfer affect the evolution of anti-biotic resistant strains of bacteria?A. It
increases the rate of beneficial mutations within a population of bacteria.
Solution
The horizontal transfer affect the evolution of anti-biotic resistant strains of bacteria, as It
increases the speed with which bacteria can adapt because they may have gotten resistance genes
from another species already.
Three main mechanism of horizontal transfer of gene occurs are transformation, transduction or
conjugation.
Transformation involves uptake of short fragments of naked DNA by naturally transformable
bacteria.
In transduction, DNA transfer occurs from one bacterium into another via bacteriophages.
In conjugation, transfer of genetic material occurs between bacterial cells by direct cell-to-cell
contact or by a bridge-like connection between two bacterial cells..
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Conjugation and transduction
- 1. KUNAL 17373016 1st YEAR IInd SEMESTER MOLECULAR GENETICS
- 2. Eg. PROKARYOTES Eg. REPRODUCTION TRANSDUCTIONCONJUGATION TRANSFORMATION
- 3. Exchange of genetic material b/w bacteria Drug resistance. Genetic material exchanged via Viruses. Drug resistance Uptake of naked DNA molecule or fragment from the medium by a cell and incorporation into recipient. Antibiotic resistance Pesticide resistance in crops
- 5. Bacterial conjugation is the transfer of genetic material between bacteria through direct cell-to-cell contact, or through a bridge-like connection between the two cells. It occurs in both gram positive and gram negative cells but the mechanism is quite different.
- 8. The transfer of genetic material from one cell to another cell by a bacteriophage is called Transduction . • Transduction has been found to occur in variety of prokaryotes , including certain species of Bacteria : Escherichia , Pseudomonas, Salmonella , Staphylococcus , etc. • The bacteriophage containing the bacterial DNA is called transduced DNA .
- 9. Generalized Transduction- Transduction in which potentially any donor bacterial gene can be transferred. Occurs during lytic cycle. Replicate Immediately. No viral DNA. Specialized Transduction - Transduction in which only certain donor genes can be transferred. Occurs during lysogenic cycle. Replicate later. Viral and bacterial DNA.