RT－PCR联合一步法扩增G蛋白α亚基基因的开放阅读框  

A COUPLED ONE-STEP METHOD OF REVERSE TRANSCRIPTION(RT)FOR GENERATION OF OPEN READING OF G PROTEIN αSUBUNIT GENE

作　　者：彭艳华[1] 黄永秀[1] 齐义鹏[1] 

机构地区：[1]武汉大学病毒系分子病毒室,中国科学院武汉植物研究所

出　　处：《武汉植物学研究》1996年第2期97-100,共4页Journal of Wuhan Botanical Research

摘　　要：报道逆转录－多聚酶链式反应（ＲＴ－ＰＣＲ）联合一步程序，扩增大于１ｋｂ的拟南芥菜Ｇ蛋白α亚基基因的开放阅读框。该程序中，当ＲＮＡ模板和引物变性，并在同一管中加入ＰＣＲ缓冲液、４种ｄＮＴＰ底物、逆转录酶和ＴａｑＤＮＡ聚合酶之后，就不需其它手工操作步骤，因而可同时进行大批量样品的操作。实验结果证明ＡＭＶ（鸟类骨髓性白血病病毒）逆转录酶对ＴａｑＤＮＡ聚合酶的活性没有抑制作用。这种ＲＴ－ＰＣＲ联合一步法可从０．５μｇ的总ＲＮＡ中快速地扩增长于１．０ｋｂ的ｃＤＮＡ片段。It is reported that a one-step reverse transcription and polymerase chain reaction (RT-PCR)procedure for amplification of open reading frame of G-protein αsubunit gene from Arabidopsis thaliana which is larger than 1 kb. In the procedure developed,manual work is minimized,larger numbers of samples can be handled. After denaturation of template RNA and the primers and addition of other regents,no further manual steps are needed, It was not found that AMV reverse transcriptase inhibited the activity of Taq-DNA polymerase,so the coupled one-step procedure of RT-PCR rapidly amplifies ORF just only from 0. 5 μg amount of total RNA.

关 键 词：拟南芥菜 RT-PCR 开放阅读框 G蛋白 Α亚基基因 

分 类 号：S634.403.2[农业科学—蔬菜学]