贵阳腐霉Pr1基因上游调控序列的克隆及分析  

Cloning and Analyzing the Upstream Sequence of the Subtilisin-like Protease Gene(Pr1) of Pythium guiyangense

作　　者：段斯亮[1] 于声[2] 苏晓庆[3] 

机构地区：[1]柳州医学高等专科学校病原生物学与免疫学教研室,广西柳州545006 [2]柳州医学高等专科学校检验教研室,广西柳州545006 [3]贵阳医学院生物学教研室,贵州贵阳550004

出　　处：《生物技术》2012年第4期5-8,共4页Biotechnology

基　　金：广西教育厅科研经费资助项目(编号:200103YB154)资助

摘　　要：目的:克隆贵阳腐霉类枯草菌素蛋白酶(Pr1)基因上游调控序列并进行分析,进一步了解Pr1基因的表达规律。方法:采用锅柄聚合酶链反应法(Panhandle PCR)扩增Pr1基因上游调控序列,并利用真核生物启动子分析软件对扩增序列进行启动子顺式作用元件预测分析。结果:获得864bp基因序列,分析表明,该序列包含2个TATA框,一个可能的转录起始区,并具备氮调控因子结合位点(相隔很近的GATA序列),碳调控因子结合位点(5’SYGGRG 3’)。这与Pr1的表达受碳/氮抑制的结果一致。结论:Panhandle PCR方法成功克隆贵阳腐霉Pr1基因上游调控序列,GenBank登录号为:JQ975036。Objective:Cloning and analyzing the upstream sequence of the Subtilisin-like Protease(Pr1)gene of Pythium guiyangense,so as to know the expression regulation of Pythium guiyangens.Method:The panhandle polymerase chain reaction(PPCR) has been used to clone the unknown sequence,which was analysed by the promoter prediction software.Result:A 864 panhandle PCR product was subsequently subcloned and the sequence analysis indicated that it contains two TATA box,potential translation initiation region.and putative binding site of nitrogen regulator protein(closely spaced GATA),putative binding site of carbon regulator protein(5’SYGGRG3’),which was in agreement with the results that Pr1 was repressed by glucose and organic nitrogen.Conclusion:Panhandle PCR was successfully cloned the upstream sequence of the Subtilisin-like Protease(Pr1)gene.The gene has been accessed by GenBank(Accession:JQ975036)

关 键 词：贵阳腐霉 锅柄聚合酶链反应 上游调控序列 类枯草菌素蛋白酶 

分 类 号：S476[农业科学—农业昆虫与害虫防治] Q78[农业科学—植物保护]