检测微囊藻毒素合成酶基因mcyH的SYBR GreenⅠ实时荧光定量RT-PCR方法的建立  

Quantification of microcystin synthetase gene mcyH using real-time quantitative RT-PCR with SYBR greenⅠ

作　　者：方志慧[1] 许敏[1] 程凯[1] 赵以军[1] 

机构地区：[1]华中师范大学生命科学学院,武汉430079

出　　处：《华中师范大学学报（自然科学版）》2008年第3期452-455,共4页Journal of Central China Normal University：Natural Sciences

基　　金：国家自然科学基金项目(30670088)

摘　　要：利用RT-PCR方法从铜绿微囊藻PCC7806 mcy基因中扩增出目的基因片段,用TA克隆方法将基因片段插入pGEMT-T Easy中来制备质粒标准品.通过不同连续稀释倍数的质粒标准品成功构建了16srRNA和mcyH两基因的标准曲线,该法构建的两基因标准曲线r2分别为0.9916和0.9938,且质粒标准品具有较好的重复性,满足实时荧光定量RT-PCR的要求.Microcystis aeruginosa 7806 mcy genes RT- PCR product was subcloned into PGEM-T vector to contrust recombinant plasmid. Standard curves of 16srRNA and mcyH were developped by different folds serial diluted dplasmid DNA, and r^2 of two standard curves separately was 0.9916 and 0. 9938 by the method, The plasmid standard has a better reproducibility. It can satisfy the experiments need.

关 键 词：mcyH 实时荧光定量RT—PCR SYBR Green Ⅰ 

分 类 号：Q756[生物学—分子生物学]