Introduction to Recombinant Genetics- Biology 350

e.coli de3 and BL21 cells

DE3 cells have a T7 RNA polymerase integrated into the host genome, usually as a lambda phage lysogen. The T7 RNA polymerase is controlled by the Lac promoter and is leaky in transcription

BL21 cells have additonal proteases that reduce the leakyness of the DE3 regulation.

scFv Expression Vector

Prokaryotic vectors

Cloning by recombination avoids finding too few or too many restriction sites

Xi-Clone

pDONOR and pDEST vectors

eukaryotic vectors

Clontech Retroviral vectors.

Baculovirus vectors.

Why use Baculovirus Expression Vector System (BECS)

Expressing more than one protein.

Promoting proper folding.