3. In this class we are using a specialized expression cell line called BL-21 pL
ID: 145706 • Letter: 3
Question
3. In this class we are using a specialized expression cell line called BL-21 pLysS produced by Agilent. This same company also produces at least two other BL-21 cell lines called ArticExpressT(DE3)competent cells and BL-21 Codon Plus®(DE3)-RIPL competent cells. Given this information answer the following questions a. If you decide to use the ArticExpressTM competent cells in your research what is the problem that you are likely encountering in your expression protocol (2 points)? b. If you decide to use the BL-21 Codon Plus®(DE3)-RIPL competent cells in your research what problem are you trying to overcome (2 points)? What additional genes do these cell-lines express (4 points)? 1. ArticExpressT (DE3): c. 2. BL-21 Codon Plus&(DE3)-RIPL: For the ArticExpressTM(DE3) cell-line what E. coli genes are these additional genes homologues of (2 points)? d. e. For the ArticExpressT(DE3) cel-line what is the primary difference between the E. coli genes and these homologues (2 points)?Explanation / Answer
a) If we decide to use the ArticExpress competent cells in our research, the problem that we are likely encountering in the expression protocol is chaperone, Cpn60 can co-purify with the target protein. And it is not possible to dissociate the target protein from chaperone.
b) If we decide to use the BL-21 codon plus-RIPL competent cells in our research, the problem we will try to overcome is as followed. As, it has a T7-based expression, it directs all gene expression in the cells to its own DNA.
c) Additional genes these cell lines express are:
1. AnticExpress (DE3): Cpn60/Cpn10, cold-adapted chaperones
2. BL-21 codon plus (DE3)-RIPL: All genes in the bacterial DNA.
d) For the ArticExpress (DE3) cell line, groEL and groES are two chaperones homologues of cpn60 and cpn10
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