About signal peptide

[Trond Erik Vee Aune]

WANG XF wrote:
> Hello, everyone!
> 
> I have a question about protein secretion. I want to develop a construct as
> follows.
> 
> *promoter----protein 1--protease site---signal peptide---protein 2*
> 
> If the two proteins was expressed and cleaved at the protease site, protein
> 2 will be secreted into the supernatant?

I assume the signal peptide is for Sec-mediated translocation to the E. 
coli periplasm. E. coli does not efficiently secrete proteins, but 
over-expression with translocation to periplasm may lead to lysis, 
addition of chemicals that disrupt the outer membrane may lead to 
secretion.

Some references:

Mergulhão, F. J. M., D. K. Summers and G. A. Monteiro (2005). 
"Recombinant protein secretion in Escherichia coli." Biotechnol. Adv. 
23(3): 177-202.
	
Mergulhão, F. J. M., M. A. Taipa, J. M. S. Cabral and G. A. Monteiro 
(2004). "Evaluation of bottlenecks in proinsulin secretion by 
Escherichia coli." J. Biotechnol. 109(1-2): 31-43.

Francetic, O., D. Belin, C. Badaut and A. P. Pugsley (2000). "Expression 
of the endogenous type II secretion pathway in Escherichia coli leads to 
chitinase secretion." EMBO. J. 19(24): 6697-6703.

Trond Erik