阴道毛滴虫AP33基因的乳酸菌表达载体的构建
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R38

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四川省教育厅课题(11ZB227);泸州市科技局课题(泸市科[2007]16号)


Construction of the expression plasmid of T.vaginalis AP33 gene in lactic acid bacteria
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    摘要:

    目的:构建阴道毛滴虫AP33基因的乳酸菌表达载体。方法:提取阴道毛滴虫总RNA为模板,经RT-PCR得到AP33基因的PCR特异性产物,将其连接入乳酸菌分泌表达载体pVE5523质粒中,并转化入TOP10菌中保存。结果 :构建的AP33-pVE5523重组表达载体,经PCR鉴定正确;获得的目的基因片段经测序与GenBank公布的阴道毛滴虫AP33基因序列100%符合。结论:成功构建AP33-pVE5523表达载体,为进一步研究阴道毛滴虫AP33基因在乳酸菌中的表达打下基础。

    Abstract:

    Objective: To construct the expression plasmid AP33-pVE5523.Methods: The total RNA extracted from the trophozoites of T.vaginalis was used as template for the amplification of AP33 gene by using RT-PCR.The amplified products were ligated with expression plasmid pVE5523 of lactic acid bacteria(LAB).Then the recombination product AP33-pVE5523 was preserved in TOP10 bacteria.Results:The recombination product AP33-pVE5523 checked by PCR was correct.The DNA sequence analysis revealed that the homology with the gene of T.vaginalis adhesion protein AP33-1 was 100%.Conclusion: The expression plasmid AP33-pVE5523 was constructed successfully,which provides basis for the reaserch of AP33 expression in LAB.

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  • 在线发布日期: 2023-08-04
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