| 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
| Cloning of β-Galactosidase Gene from Lactobacillus acidophilus and Study of Its Activity as a Food-Grade Selection Marker
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| DOI：
| 中文关键词: β-半乳糖苷酶 嗜酸乳杆菌 乳酸乳球菌 食品级筛选标记
| 英文关键词: β-galactosidase Lactobacillus acidophilus Lactococcus lactis food-grade selection marker
| 基金项目:重庆市教委资助项目(JK060306)
| 作者
| 单位
| 贺松
| 重庆医科大学,分子医学与肿瘤研究中心,重庆,400016
| 龚芳红
| 重庆医科大学,分子医学与肿瘤研究中心,重庆,400016
| 张德纯
| 重庆医科大学,分子医学与肿瘤研究中心,重庆,400016
| 刘明方
| 重庆医科大学,分子医学与肿瘤研究中心,重庆,400016
| 程芳
| 西安交通大学,医学院,陕西,西安,710061
| 郭亚楠
| 重庆医科大学,分子医学与肿瘤研究中心,重庆,400016
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| 中文摘要:
| 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。
| 英文摘要:
| 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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