PPT 2

Question 1

Yersinia pseudotuberculosis

Answer 1

enteropathogen transmitted via contaminated food
-produces Far East Scarlet Fever
-genetic ancestor of Y. pestis
-adhesion is a critical step in pathogenesis
(fever and right sided abdominal pain)

Question 2

How do we identify adhesins from Y. pseudotuberculosis?

Answer 2

Complementation: isolate the DNA and cut, splice the donor DNA into plasmid, introduce into recipient, enrich for invasive colonies, use sequence DNA by identifidying using SDS PAGE or Mutate and reintroduce into TB

Question 3

Transpon Mutagenesis

Answer 3

transpons are small pieces of DNA that mobilize themselves to another place in the genome

• bind to a target sequence and insert DNA at that site (different ones)
• used to increase library of mutants
• determine resistant genes
• appearance and growth ability

Question 4

Tn-SEQ analysis

Answer 4

more parallel than signature-tagged mutagenesis
10-100k mutants screened
-sequencing/counting specific chromosomal location using parallel sequencing