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