№ 5 / 2014 / 37-42

Cloning and analysis of expression of synthetic human apolipoprotein А-I genes in Escherichia coli and methylotrophic yeasts Pichia pastoris

Abstract

The expression of synthetic genes of human ApoA-I protein in the methylotrophic yeasts Pichia pastoris X-33 and E. coli Rosetta 2(DE3) cells were studied. It is shown that expression of the ApoA-I genes in yeast cells is accompanied by cell lysis and accumulation of unprocessed ApoA-I protein in the growth medium, which is probably due to the amphiphilic properties of the protein. Expression of ApoA-I gene in E. coli Rosetta ™ (DE3) cells provides the protein of interest with the yield of 50 mg/l. Recombinant polypeptide reacts with antibodies against native human ApoAI and has close molecular weight as native protein.

Key words

E. coli Rosetta 2(DE3), Pichia pastoris X-33, apolipoprotein А-I, affinity purification, сloning, recombinant protein, gene expression

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About Authors (Correspondence):

Mamaev A.L. – researcher of the genetic engineering laboratory, e-mail: alexeymamaev@inbox.ru

Beklemishev A.B. – doctor of biological sciences, professor, head of the genetic engineering laboratory, e-mail: beklem@ niibch.ru

Full Text

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Received: 08/02/2015