Microbial Genetics

Serological classification -> Analysis of microbes in terms of their phenotype and

metabolic by-products

Genomic classification -> Analysis of microbes in terms of their genetic material

Central Dogma of Genetics

- DNA-dependent DNA Polymerase replicates and copies DNA
- DNA-dependent RNA Polymerase transcribes DNA in terms of mRNA
- Ribosomes converts mRNA into proteins with the aid of aminoacyl-tRNAs
- Positive sense RNA -> 3-5
- Negative sense RNA -> 5-3
- Ribosomes can only recognize single-stranded RNA
- Expression of genes lead to protein production
- RNA-dependent RNA Polymerase converts RNA into double stranded RNA
- Totipotency ->
- Transgenesis -> Transfer of genes from 1 organism to another
- pGLO plasmid -> plasmid vector used by Bio-RAD to transfer gene via
horizontal gene transfer
o ori region -> site where replication of plasmid begins
o bla region -> gene that codes for beta-lactamase, which inactivates
penicillum
o pBAD promoter -> site where transcription of the plasmid begins
binding of pBAD to RNA Polymerase promotes transcription of
the pGLO plasmid
o araC operon -> gene that codes for arabinose-binding protein which
makes RNA polymerase bind to the pBAD promoter
o GFP gene -> gene that codes for green fluorescent protein, expression
of this gene shows that the plasmid was accepted and can be
expressed by a certain organism
o Restriction enzymes
Cut DNA specifically into fragments
EcoR1 is a restriction enzyme that originated from E.coli
HindIII is a restriction enzyme that originated from Hemophilus
Influenzae
- Plasmid + Gene of Interest -> Insertion to bacteria -> Expression
o Bacteria may not take up the plasmid if it does not deem it necessary
o Bacteria should be placed in an environment wherein it could only
survive it it took the plasmid containing the gene of interest
 Plasmid should contain Beta-lactamase gene to ensure that it
can survive the environment (environment which contains Beta-
lactam environment)
o GFP should still be present in the control plasmid
Expression of the GFP by the bacteria indicates that the plasmid
containing the gene of interest was accepted and is being
expressed by the bacteria
o Messelson-Stahl experiment
Showed that DNA replication is semiconservative (one strand of
DNA from both parents is replicated to produce the daughter
DNA)
Parent strands were tagged with 15N and allowed to replicate
and separated using CsCl gradient centrifugation
o Proteins of DNA Replication
Topoisomerase
Single-strand binding proteins
Stabilizes the single stranded unwinded DNA and
prevents it from being hydrolyzed
DNA Polymerase