High-level expression of human proapolipoprotein A-I in Escherichia coli using expression plasmids containing tandemly polymerized proapolipoprotein A-I structural genes
A gene coding human proapolipoprotein A-I (proapo A-I) was inserted into a plasmid with a consensus ribosome binding sequence in Escherichia coli. One to three copies of this gene were tandemly inserted to construct proapo A-I expression plasmids, pMTpAI, pMT(pAI)2 and pMT(pAI)3, respectively. The cells harbouring pMT(pAI)3, could produce proapo A-I at a level of 49 μg/ml A600 and up to approx. 48% of the total cellular protein. The product was soluble in E. coli, and formed protein-lipid complexes with dimyristoyl phosphatidyl choline, which formed stacked disc structures. Analyses of the rec proapo A-I formed in the bacteria was identical to human proapo A-I except for methionine at the N-terminus.
KeywordsEscherichia Coli Methionine Choline Structural Gene Phosphatidyl Choline
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