| 以thyA基因为选择压力非抗性质粒载体的构建 |
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| 引用本文： | 黄瑜,何召庆,张莉,刘尚高,王春,秦泽荣,孙哲. 以thyA基因为选择压力非抗性质粒载体的构建[J]. 微生物学通报, 2001, 28(2): 42-46 |
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| 作者姓名： | 黄瑜 何召庆 张莉 刘尚高 王春 秦泽荣 孙哲 |
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| 作者单位： | 1. 中国农业大学动物医学院
2. 中国农业大学动物医学院 吉林农业大学动物科技学院 |
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| 基金项目： | 国家自然科学基金资助项目(No.39980017)Projection Granted by Chinese National Natural Science Fund(No. 39980017) |
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| 摘 要： | 以干酷乳杆菌L.casei34103染色体DNA为模板，利用PCR技术扩增胸苷酸合成酶（Thymidylatesynthase,thyA)基因，回收纯化，选择以红霉素抗性为选择压力的可以在大肠杆菌和乳酸菌中穿梭表达的质粒pW425e为基本质粒，以thyA基因取代红霉素基因，获得重组载体并鉴定，此重组载体可以对thyA基因缺陷的大肠杆菌E.coli X51和嗜酸乳杆菌DOMLaS 107进行功能弥补，进而构建了以thyA基因为地选择压力的非抗生素抗性穿梭表达载体，其大小为3716bp，并命名为pW425t。
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| 关 键 词： | thyA基因 选择压力 穿梭载体 构建 质粒 |
| 文章编号： | 0253-2654(2001)02-0042-05 |
| 修稿时间： | 1999-11-12 |
CONSTRUCTION OF SHUTTLE VECTOR BY USING THYA GENE AS A SELECTIVE PRESSURE |
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| Abstract: | Using chromosome DNA of Lactobacillus casei 34103 as template, thy A (Thymidylate synthase) gene was amplified by PCR with pfu DNA polymerase. Choosing pW425e vector, which can express shuttlely between E. coli and Lactobacillus,containing erythromycin resistance gene, as the basic plasmid. The PCR production of thyA gene was used to replace the erythromycin resistance gene of pW425e. The new plasmid vector consits of 3716bp as expected. Which can remedy the thyA mutant E. coli X51 and Lactobacillus DOMLaS107 successfully, and named as pW425t. |
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| Keywords: | thyA gene Selective pressure Shuttle vector Construction |
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