Question

1-Give two characteristics of recombinant proteins cloned into the pRSET-FP plasmid.

2-Briefly explain what the Ori locus is and its purpose?

3-What is the selective media? What is it selecting for (briefly explain)?

Answer 1

Ans 1. Recombinant proteins cloned into pRSET-FP plasmid

1. Helps in overexpression of proteins as it has T7 promoter and can be regulated by IPTG induction.

2. Also vector allows to visualise and localise the expressed protein in the cell as it has cyan fluoroscent protein tags gene coding sequence.

Ans 2. Ori locus is the origin of a plasmid at which DNA replication will be initiated. At this point hydrogen bonds between two strands of DNA will start melting (breaking) by enzymes of the host. It is basically a nucleotide sequence which is recognised by enzymes involved in DNA replication of a plasmid DNA.

Ans 3. Selective media is the microbial growth media that contains nutrients for the growth of bacteria having certain properties while inhibiting the growth of other microorganisms in a mixed culture.

for example, one has to isolate the gram negative bacteria from a mixed population of bacteria, one can use media containing main essential nutrients required for growth as well as other substances like dyes (eosin methylene blue) which will allow the growth of gram negative bacteria but not that of other bacterial population.

This helps in isolating pure culture of bacterial cells showing similar properties.