关键词: 嗜酸乳杆菌, 亚油酸异构酶, 基因克隆, 重组蛋白, 纯化

Abstract:

This paper deals with the cloning and expression of conjugated linoleic acid isomerase gene of Lactobaciι lus αcidophilus AS 1. 1854 by means of molecular biologïcal method. In the investigation , first , the genome was ex-tracted after the cel1 was cultured and gathered , and was amplified by means of PCR. Next , the amplified gene was cloned into pET30a plasmid vector and subsequently converted into Escherichia coli BL21 strain. Then , the gene was induced to express the recombinant protein at a low temperature , and the recombinant protein was separated and purified with Ni-NTA agarose. Finally , the target gene was amplified and converted into plasmid , and the re-combinant protein was expressed and separated through the experiment. It is found that soluble recombinant conju-gated linoleic acid isomerase can be expressed in prokaryotic cell, and that the target protein can be obtained by washing wÎth 200mmol/L imidazole.

Key words: Lactobacillus acidophilus, conjugated linoleic acid isomerase, gene cloning, recombinant protein, pu-rification