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Transcript
Horizontal Gene Transfer: New Gene Acquisition Transformation: naked DNA uptake by bacteria Transduction: bacterial DNA transferred by viruses (phage) Conjugation: DNA transfer between bacterial cells Furuya EY and Lowy F (2006) Nat Rev Microbiol. 4: 36–45. Horizontal Gene Transfer: New Gene Acquisition – Selfish mechanisms result in genetic diversity • Often confer properties of virulence, antibiotic resistance, or metabolic fitness – May provide different characteristics • • • • Utilization of energy sources Acid tolerance Development of symbiosis Ability to cause disease – Pathogenicity islands Horizontal Gene Transfer: New Gene Acquisition 1. Lyse antibiotic resistant cells SDS + 60˚C Why are we using DNA that has an antibiotic resistant marker? How are we able to lyse the cells? DNA component with antibiotic resistance marker gets released Horizontal Gene Transfer: New Gene Acquisition 2. Combine free DNA with cells sensitive to the antibiotic and plate on non-selective media + Horizontal Gene Transfer: New Gene Acquisition 3. Plate transformed cells on plates selective for antibiotic resistance Horizontal Gene Transfer: New Gene Acquisition a) DNA only: test that it is inert/not-alive b) StrS cells c) StrR cells d) 1 loopful StrS cells. Place 1 loopful DNA ON TOP of those cells e) 1 loopful StrS cells. THEN one loopful DNase THEN one loopful DNA b) StrS cells c) StrR cells d) StrS cells + DNA a) DNA only e) StrS cells + DNase + DNA