满江红鱼腥藻glnA基因上游调控区的克隆及其序列分析  

Cloning and Nucleotide Sequence of Leader Region of glnA from Aac

作　　者：高志环[1] 刘祥林[1] 印莉萍[1] 吴晓强[1] 邱泽生[1] 

机构地区：[1]首都师范大学生物系

出　　处：《北京师范学院学报（自然科学版）》1998年第2期54-58,共5页

基　　金：国家自然科学基金

摘　　要：以满江红鱼腥藻（Ａａｃ）提取总ＤＮＡ为模板，通过ＰＣＲ技术，扩增得到其谷氨酰胺合成酶基因（ｇｌｎＡ）上游区．经酶切得到４０９ｂｐ的片段，并将其连接到ｐＢｌｕｅｓｃｒｉｐｔⅡｋｓ＋上测序．将测序结果与Ａｎａｂａｅｎａ７１２０调控序列比较，有９８２％的同源性．在上游调控中也具有ｎｉｆｌｉｋｅ和Ｅ．ｃｏｌｉｌｉｋｅ启动子，值得注意的是，调节蛋白ＶＦ１的结合位点正好位于ｎｉｆｌｉｋｅ启动子上．所克隆的片段不但对蓝藻固氮、泌氨的基因调控研究具有意义，也对表达载体的构建具有实用价值．The total DNA derived from Anabeana azolla(Aac) was used as template and the up stream of glnA gene was obtained by PCR.The derived 409bp element was linked to pBluescriptⅡ KS + plasmid after digested with restriction endonulease.The sequence result indicated that this glnA gene promoter has 98 2% homology with the one obtained from Anabaena 7120.In addition,it contained the E.coli like and nif like promoters in up stream.Noteworthly,the binding site of regulatory protein VF1 is right on nif like promoter.This cloned fragment is valuable for not only the research of regulatory genes in the N 2 fixing and secreting NH + 4 of Arabeana azolloa but also for the construction of expressive vectors.

关 键 词：满江红鱼腥藻 AAC glnA基因 上游调控区 克隆 序列分析 PCR技术 谷氨酰胺合成酶 GS 总DNA 结合位点 

分 类 号：Q949.22[生物学—植物学] Q78