玉米ZmPsaH基因真核表达载体的构建及转化

Construction and Transformation of Eukaryotic Express Vector of ZmPsaH Gene from Maize

【目的】PsaH是光合系统Ⅰ(PSI)中的一个亚基,在植物光合系统中起着重要的作用。为研究玉米ZmPsaH基因在抗逆中的作用提供基础。【方法】构建玉米ZmPsaH基因真核表达载体是了解玉米中该基因功能的重要途径之一。利用RT-PCR技术扩增得到大小约为420 bp的玉米ZmPsaH基因,经限制性内切酶SalI和NotI进行消化,将目的基因与真核表达质粒pREP-5N连接,获得重组真核表达质粒ZmPsaH-5N。【结果】通过菌落PCR、双酶切及测序等方法进行鉴定,重组真核表达质粒ZmPsaH-5N构建成功。制备酵母感受态,将重组质粒电击转入酵母中,酵母菌落PCR结果显示电击转化成功。【结论】成功构建了玉米ZmPsaH基因真核表达载体,并且成功转化了酵母。

[Objective] The psaH gene is one subunit of photosystem Ⅰ,and it plays an important role in Photosynthesis.The purpose of this paper is to provide the basis for the study of ZmpsaH gene on resistance.[Method] Construction of ZmPsaH eukaryotic expression vector is an important way to understand the gene function in maize.The ZmPsaH gene was cloned from the maize by RT-PCR,which was named ZmPsaH （photosystem Ⅰ reaction center subunit H）.It was digested by the restriction enzymes SalⅠ and NotⅠ,and then connected with pREP-5N to obtain the recombinant eukaryotic expression plasmid ZmPsaH-5N.[Result]Identified by colony PCR,restriction enzyme digestion and sequencing methods,the results showed that the recombinant eukaryotic expression the plasmid ZmPsaH-5N was successfully constructed.Preparation of yeast competent,electric shock,the recombinant plasmid was transformed into yeast.A yeast drop-PCR result showed that electroporation was successful.[Conclusion] Construction of ZmPsaH gene eukaryotic expression vector and transformation are successful.