嗜酸乳杆菌β-半乳糖苷酶基因的克隆及其作为食品级筛选标记的研究

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嗜酸乳杆菌β-半乳糖苷酶基因的克隆及其作为食品级筛选标记的研究
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基金项目:重庆市教委资助项目(JK060306)

Cloning of β-Galactosidase Gene from Lactobacillus acidophilus and Study of Its Activity as a Food-Grade Selection Marker

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为研究嗜酸乳杆菌β-半乳糖苷酶基因(lacZ)作为食品级筛选标记的活性及筛选稳定性,作者从嗜酸乳杆菌基因组中克隆β-半乳糖苷酶基因并在乳酸乳球菌中表达,通过X-gal显色筛选阳性克隆;用X-gal显色筛选方法将重组乳酸乳球菌传60代,并测定β-半乳糖苷酶酶活和比活力。结果显示:通过表达有活性的β-半乳糖苷酶和X-gal显色可成功筛选出阳性克隆菌株;对借助X-gal传60代后的重组乳酸乳球菌进行β-半乳糖苷酶比活力检测,与X-gal筛选的第二代相比没有显著差异(P=0.592>0.05),与红霉素筛选比较也没有显著差异(P=0.882>0.05)。由此可见,β-半乳糖苷酶基因作为筛选标记具有较好的活性和稳定性,为以β-半乳糖苷酶基因作为筛选标记的食品级载体的研究奠定了基础。

Abstract:

To study the selective activity and stability of β-galactosidase gene(lacZ) from Lactobacillus acidophilus as a food-grade selection marker,the lacZ gene from L.acidophilus was cloned and expressed in L.lactis.In this study,the selective activity was determined by X-gal-containing medium and the activity and specific activity of β-galactosidase by X-gal.The results showed that the positive recombinant strains were selected successfully by β-galactosidase activity and X-gal coloration.The β-galactosidase specific activity of the recombinant L.lactis after a period of 60 generations selected with X-gal was similar to that of the 2nd generation(P=0.592>0.05),and also similar to the 61st generation selected with Erythromycin(P=0.882>0.05).These results indicated that the β-galactosidase lacZ gene had good activity and stability as a selection marker,and it could be used as a selection marker for construction of a food-grade vector.

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贺松,龚芳红,张德纯,刘明方,程芳,郭亚楠.嗜酸乳杆菌β-半乳糖苷酶基因的克隆及其作为食品级筛选标记的研究[J].食品与生物技术学报,2010,29(5):777-781.

HE Song, GONG Fang-hong, ZHANG De-chun, LIU Ming-fang, CHENG Fang, GUO Ya-nan. Cloning of β-Galactosidase Gene from Lactobacillus acidophilus and Study of Its Activity as a Food-Grade Selection Marker[J]. Journal of Food Science and Biotechnology,2010,29(5):777-781.

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在线发布日期: 2014-06-17
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