M.hydrocarbonoclasticus NY4中nosZ基因的克隆与异源表达
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国家自然科学基金项目(31270162);江苏高校品牌专业建设工程资助项目(TAPP)


Cloning and heterologous expression of nosZ gene in Marinobacter hydrocarbonoclasticus NY4
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    摘要:

    选取反硝化菌株Marinobacter hydrocarbonoclasticus NY4为试验对象,首先通过反硝化气态产物分析,证实了该菌株中N2OR的存在;随后设计特异性引物并通过PCR扩增获取了nosZ的全长基因(1 896 bp),该基因与来源于Geobacillus thermodenitrificans的nosZ基因仅保持了46.9%的相似性;实现了nosZ基因在大肠杆菌中的异源表达,并成功检测到了N2OR酶活,其比活力达到(0.2±0.01) U/mg。

    Abstract:

    The nosZ gene encoded N2OR was cloned and expressed heterologously.One denitrifying strain named as Marinobacter hydrocarbonoclasticus NY4 was used in the experiment.The existence of N2OR in strain NY4 was confirmed by analyzing gaseous products produced by denitrification.The full-length nosZ gene (1 896 bp) was obtained by PCR amplification with a pairs of specific primers,which only shared 46.9% similarity with the nosZ gene from G. thermodenitrificans. The heterologous expression of the nosZ gene in Escherichia coli was carried out.The nosZ gene from strain NY4 was further expressed in E. coli.The activity of N2OR was successfully detected.The enzyme activity was (0.20±0.01) U/mg.

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杨立志,王鹏胤,胡南. M. hydrocarbonoclasticus NY4中nosZ基因的克隆与异源表达[J].华中农业大学学报,2017,36(2):64-68

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  • 收稿日期:2016-04-19
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  • 在线发布日期: 2017-02-27
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