| 嗜热链球菌胸苷酸合成酶基因的克隆与序列分析 |
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| 引用本文： | 张艳禾,朱晓俊,张兰威.嗜热链球菌胸苷酸合成酶基因的克隆与序列分析[J].食品科学,2007,28(4):223-227. |
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| 作者姓名： | 张艳禾 朱晓俊 张兰威 |
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| 作者单位： | 东北农业大学食品学院； 黑龙江生物制品一厂； 东北农业大学食品学院 黑龙江 哈尔滨 150030； 黑龙江 哈尔滨 150069； 黑龙江 哈尔滨 150030 哈尔滨工业大学食品科学与工程学院； 黑龙江 哈尔滨 150001； |
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| 摘 要： | 本研究以嗜热链球菌(S．thermophilus)染色体DNA为模板,利用PCR技术扩增出胸苷酸合成酶(thymidylate synthase,thyA)基因,将其克隆入T载体中,转化DH5α,筛选阳性克隆,提取质粒,进行酶切鉴定,PCR扩增鉴定,进行测序,与已知序列进行同源性比较,结果表明成功克隆了thyA基因,全长900bp,与国外报道的S．thermophilus ATCC19258 thyA基因同源性达99．9%。这为构建以thyA基因为选择压力的非抗生素抗性载体提供了理论依据。
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| 关 键 词： | thyA基因 克隆 序列分析 |
| 文章编号： | 1002-6630（2007）04-0223-05 |
| 修稿时间： | 2006-04-30 |
Clone and Sequence Analysis of S. thermophilus Thymidylate Synthase Gene |
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| ZHANG Yan-he,ZHU Xiao-jun,ZHANG Lan-wei.Clone and Sequence Analysis of S. thermophilus Thymidylate Synthase Gene[J].Food Science,2007,28(4):223-227. |
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| Authors: | ZHANG Yan-he ZHU Xiao-jun ZHANG Lan-wei |
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| Affiliation: | 1.College of Food Science,Northeast Agricultural University,Harbin 150030,China; 2.First Bioproducts Manufactory of Heilongjiang, Harbin 150069,China; 3.School of Food Science and Engineering,Harbin Institute of Technology,Harbin 150001,China |
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| Abstract: | Using chromosome DNA of S.thermophilus as template.Thymidylate sythase(thyA)gene was amplified by PCR with ExTaq DNA polymerase,and purified by purification kit.Subsequently,the PCR production of thyA gene was cloned into the pMD-18T,transformed into DH5αThe recombination plasmid containing thyA gene was selected and identified by restriction enzyme analysis and by PCR amplification and sequence.The results showed that the whole thyA gene is cloned already(about 900bp).Sequence analysis showed that the thyA gene displays 99.9% nucleotide identities with the published sequences.The thyA gene offers a good foundation for the shuttle vector construction by thymidylate synthase(thyA)gene as a selective pressure. |
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| Keywords: | thyA gene cloning sequence analysis |
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