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９８２％的同源性．在上游调控中也具有ｎｉｆｌｉｋｅ和Ｅ．ｃｏｌｉｌｉｋｅ启动子，值得注意的是，调节蛋白ＶＦ１的结合位点正好位于ｎｉｆｌｉｋｅ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.
基金
国家自然科学基金
