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Bacterial Conjugation

The document discusses genetic transfer between prokaryotes through horizontal gene transfer mechanisms such as transformation, transduction, and conjugation. It specifically focuses on bacterial conjugation, describing the process where an F+ donor bacterium transfers genetic material to an F- recipient bacterium through direct cell contact via pilus formation. The transferred DNA can contain traits for antibiotic resistance, allowing the recipient to also become resistant. An experiment is described where conjugation between an HFR donor and F- recipient on selective media allows for the selection and growth of recombinant cells.

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Batool Ashraf
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83 views

Bacterial Conjugation

The document discusses genetic transfer between prokaryotes through horizontal gene transfer mechanisms such as transformation, transduction, and conjugation. It specifically focuses on bacterial conjugation, describing the process where an F+ donor bacterium transfers genetic material to an F- recipient bacterium through direct cell contact via pilus formation. The transferred DNA can contain traits for antibiotic resistance, allowing the recipient to also become resistant. An experiment is described where conjugation between an HFR donor and F- recipient on selective media allows for the selection and growth of recombinant cells.

Uploaded by

Batool Ashraf
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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 

Richa Banthia Ben Anderson


 Genetic variability – ESSENTIAL!!!
o Survival of the species
 Diploid Eukaryotes
o Crossing over
o Meiosis
 Haploid Prokaryotes
o Transduction
o Transformation
o Conjugation
 Genetic material exchanged via Viruses
o E.g. bacteriophage lambda
 No physical contact b/w bacteria
 Promotes drug resistance
 Horizontal gene transfer
 Acquisition
of exogenous DNA fragments from
environment
o E.g. Plasmids
 Antibioticresistance
 Pesticide resistance in crops
o E.g. Agrobacterium tumefaciens
 Horizontal gene transfer
 Exchange of genetic material b/w bacteria
 Physical contact required
 DNA exchanged via Pilli
 Drug resistance!!
 Horizontal Gene Transfer
 
Overview
The Donor The Recipient The Conjugate
 F+  F-  HFR
o Fertility factor o No fertility o High
 Contains
factor frequency
 Receives
recombinant
genetic
material for genetic  Resulting

gene material genetic


exchange  “Female” cell
recombinant
 “male” cell  Transfers
entire genomic
DNA when
Attachment Via Pilli

 Formation of “sex pilus” or F-bridge between


cells
Genetic Transfer of ssDNA

 TargetDNA is replicated in F+
 Genetic transfer of ssDNA to F-
Synthesis of
Complementary DNA

 Complementary strands of DNA are


replicated in both F- and F+
 Target DNA is incorporated into F- genome
Separation and Formation
of Two F+ cells

 Cells separate
 Form two F+ cells with completed recombinant plasmids
 Both serve as donor cells for further conjugation
 
Experiment 3
HFR (F+) F- Mixed Culture
 Able to produce  Requires amino  Able to produce
amino acids acids: Thr and Leu
o Thr, Leu, Thi o Thr, Leu, Thi o Thi not produced

 Sensitive to  Resistant to  Resistant to


Streptomycin Streptomycin Strepromycin
 HFR strain used
for experimental
purposes
M9ST Agar
 Minimal Media 9
 Streptomycin
o Antibiotic formation
o Inhibition of F- growth
 Thiamine
o Inhibition of F+ growth
o Promotion of HFR
growth
 Mix F- and HFR (F+) strains
 Incubation in 37o water bath for proper
conjugation
o 30 minutes*
 Vortex to terminate genetic transfer
 Centrifugation – DNA Pellet formation
o Removal of supernatant (TSB broth)*
 Resuspension with M9 Broth
 Plated on M9ST Agar plates
 Incubation at 37o for 48hrs
HFR (F+) F- Mixed Culture
 No growth  No Growth  Growth

 Sensitivity to  Absence of  Produced Thr

Antibiotics amino acids and Leu


 Antibiotic
Resistance

HF F-
R
http://www.youtube.com/watch?v=O-EdX4MaMFE
 http://www.sci.sdsu.edu/~smaloy/MicrobialGenetics/topic
s/plasmids/conjugation-mech.html
 BIOL 3116 Advanced Microbiology Lab Manual

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