β半乳糖苷酶基因工程乳酸菌的构建及其应用研究营养与食品卫生学专业论文.docxVIP

的方法，在国内外尚未见相似的报道。Caco．2细胞是一种适合 的方法，在国内外尚未见相似的报道。Caco．2细胞是一种适合 于研究细菌与细胞间相互作用及细胞毒性的较好的体外细胞模 型。本研究采用此细胞模型，结果表明工程菌对乳糖引起的细 胞毒具有缓解作用。 ■ 关键词B-半乳糖苷酶／质敞乳酸菌，乳球菌i乳杆菌；基因工程乳 酸菌， Studies Studies on Construction and Expression of 13 一galactosidase genetic engineering Lactococcus lactis Tutor Zhang Caowu Professor Postgraduate Lu Xiaoying Abstract objectlve：Expression of 13-galactosidase gene from balgaria Bacillus in the Lactococcus lactis host·vector system．and reconstruct this engineering system imo a food grade secretory expression system which is utilized in the field of food，medicine and drug． Methods：1)、Authentication of the Lactococcus lactis host．vector system；2)、Authentication of the Bulgaria Lactobacillus from which B -galactosidase gene comes from and screen of the bacterium which produces large quantity of且·galactosidase；3)、Construction of B —galactosidase fusion protein gene engineering Lactococcus lactis and the authentication of it；4)、Expression of 0-galactosidaSe gene by using Shine—Dalgarno sequence in plasmid pMG36e vector；51、Caco-2 cell was adopted to study the toxin effect of the genetic engineering Lactococcus lactis and the genetic engineering E．coli．Also，Caco-2 cell model was 4 used used to study the relief effect of these two genetic engineering bacteria on the cell toxin caused by lactose． Results：1)、The plasmid pMG36e used in this study has anti- erythromycin character,and subspeciece ofthe Lactococcus lactis MGl363 is subsp．cremoris．while that ofLactococcus lactis IL 1403 is subsp．Lactis； 2)、Bulgaria Lactobacillus 1．1408 produces large quantity of 13 一galactosidase；3)、With 13 amino acids from the vector in the N terminal B．galactosidase fusion protein that still has the enzyme activity is successfully expressed both in the Lactococcus lactic MGl363 and in the Lactococcus lactic ILl403；4)、The SD sequence downstream the promoter ofthe vector call be successfully recognized by either the host Lactococcus lactis MGl363 or the ILl403，and the B-galacto