BIO 326R 1st Edition Lecture 14 Outline of Last Lecture I. Cell Division- Binary FissionII. Cell biology of bacteriaa. Green fluorescent proteinb. FtsZc. MinC, MinD, MinEd. MreBe. Dividing on multiple planesOutline of Current Lecture I. Phenotype vs Genotypea. MutantII. Plasmidsa. Namingb. Host rangec. Replicating plasmidd. Plasmid copy numberi. RNA I, RNA II, RopCurrent LecturePhenotype vs. Genotype- Phenotype= observable traits—expressed genes- Genotype= DNA sequenceo Ex: E. coli can produce the amino acid argenine and a mutant has a deletion in the protein argB, used to synthesize argenine Genotype= deletion argB Phenotype= argenine auxotroph (=required for growth)- Mutant= strain that has a mutation genotypic change that usually results in a phenotypeo From example: deletion argB= arginine mutanto Clicker Q: if minC is a mutant, ring will form but not at the mid-cell, because minCfinds the mid-cellPlasmids- Plasmid= autonomous (replicate independently from chromosome) replicating DNA molecules (replicons)o Host machinery required for replication Use DNA polymerase I and III and DNA ligase from the hosto Most are circular, but some are linear- Naming a plasmidThese notes represent a detailed interpretation of the professor’s lecture. GradeBuddy is best used as a supplement to your own notes, not as a substitute.o Ex: pLZ200 Lower case p = plasmid LZ= initials of who made the plasmid 200= a number you choose- Host range=> not all plasmids replicate in all bacteriao Host= bacteriao Range= which bacteria/ how many the plasmid will replicate ino Narrow host range= very few bacteriao Broad range= many different bacteria Encode additional factors to replicate in many different bacteria Encode many factors that help find the origins of replication in different types of bacteria- Replication of the plasmido Theta= like replicating the chromosomeo Rolling circle= conjugation Double stranded circular DNA, nick on the outer strand, newly synthesizesin place of nick and around the outer circle—now a new outer strand, “old” strand from outer circle comes off and copies to make a new plasmid- Copy numbero pMB1 Col E1 maintained at 15-20 copies/cello how do we maintain this copy number? RNA II is processed (cleaved) by a host protein called RNAse H can initiate replication RNA I is antisense to RNA II- I can bind to II and prevent RNA II from initiating replication Rop helps RNA I bind to RNA II RNA II: RNA I ratio allows for 15-20 copy number- More RNA II than RNA
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