玉米大斑病菌黑色素合成还原酶基因酵母单杂交报告载体的构建  

Construction of Report Vectors of Melanin HNreductase Gene of Setosphaeria turcica in Yeast One-hybrid

作　　者：贾慧[1] 郭丽婕[1] 孟庆江[1] 曹志艳[1] 宋亚凤 

机构地区：[1]河北农业大学生命科学学院/河北省植物生理与分子病理学重点实验室,河北保定071000

出　　处：《湖北农业科学》2015年第14期3542-3545,共4页Hubei Agricultural Sciences

基　　金：河北省自然科学基金项目(C2011204066)

摘　　要：根据玉米大斑病菌(Setosphaeria turcica)黑色素合成还原酶基因St3HNR、St4HNR 5′端侧翼序列和酵母单杂交报告载体p HIS2.1多克隆位点,设计了带有限制性内切酶位点的特异性引物,并以玉米大斑病菌01-23菌株基因组DNA为模板,PCR扩增启动子区域长度约1 000 bp的片段,双酶切目的片段及载体p HIS2.1回收并连接,构建2个还原酶基因的酵母单杂交报告载体。结果表明,克隆到启动子区域长度为998 bp核酸片段,经PCR检测、测序、酶切鉴定,2个还原酶基因的酵母单杂交报告载体构建成功,为进一步研究还原酶与转录因子互作关系及明确黑素素合成调控机制奠定了基础。According to the restriction enzyme sites of melanin biosynthesis promoter region of reductase gene St3 HNR,St4HNR and reporter vector pHIS2.1 of Setosphaeria turcica, two pairs of primers containing restriction enzyme sites were designed, the fragment length about 1 000 bp obtained by PCR using genomic DNA of S.turcica 01-23 strain as templet, and double digested by the corresponding restricted enzymes respectively, recycled and connected, constructed the yeast one-hybrid reporter vector of reductase gene. The results showed that the fragment length of two genes were both 998 bp, vectors were successfully constructed by PCR, sequencing, restriction enzyme digestion, and lay the foundation for the study of the transcription factor.

关 键 词：玉米大斑病菌(Setosphaeria turcica) 黑色素 酵母单杂交报告载体 

分 类 号：Q78[生物学—分子生物学]