Question

what is the fuction of these follwoing in Vector?

CMV promoter

SV 40 poly A

SV 40 origion

hGH poly A

f1 origion

Amp

neo

pUC

pBR322

kan

gene X

T7 promter

6xHis

LacO

ColE1 ori

M13 ori

Answer 1

CMV promoter: It is a strong mammalian expression promoter help in gene expression.

SV 40 poly A: It enhance protein production in baculovirus expression vector system. The signal is recognized by host RNA polymerase II complex to process precursor mRNA and increase the stability of the mature mRNA. It also enhance the efficiency of mRNA translation in eukaryotic cells.

SV 40 origin: SV40 origin of replication allows autonomous (as an episome) replication in mammalian cells expressing the SV40 large T-antigen, They are mainly used in the shuttle vctor.

hGH poly A: One of the most commonly used mamalian terminator having the sequence motif AAUAAA which promotes both polyadenylation and termination.

f1 origin: f1 ori enables single stranded DNA replication and packaging into phage particles.

Amp: It gives the vector Ampicillin antibiotic resistance property.

neo: It gives the vector Neomycin antibiotic resistance property

pUC: It is a type of bacterial origin of replication which helps to produce large quantities of DNA

pBR322: It is also an example of origin of replication. Infact pUC origin derived from pBR322 after two mutations.

kan: It gives the vector Kanamycin antiobiotic resistance property.

T7 promoter: It enables the vector to allow transcription of RNA using T7 RNA polymerase. It controls the binding of the RNA polymerase and transcription factors.

6X His: It makes the expressed protein histidine tagged, which help in protein purification.

LacO: Allow regulated and inducible protein expression in E.coli

ColE1 ori: It acts as an origin of replication

M13ori: It also acts as an origin of replication. It helps in single stranded DNA replication.