摘要： 根据Taq酶既有聚合酶活性又有反转录酶活性的特点,探索了只在Taq酶单独作用下完成RT PCR反应的条件,以达到对马铃薯纺锤块茎类病毒(PSTV)的快速检测。结果表明,反转录阶段退火温度在37℃、循环数不少于10次的情况下,PCR扩增能得到一条相关特异带。经过嵌合式PCR证明,这条带为PSTV的特异带。与其他检测方法相比,用单酶法RT PCR检测PSTV步骤简单,成本低廉。
关键词:
马铃薯纺锤块茎类病毒,
单酶法RTPCR,
Taq酶
Abstract: In single-enzyme RT-PCR,Taq enzyme plays,roles of both reverse-transcripise and DNA polymerise.The reaction conditions of single-enzyme RT-PCR was researched in this paper in order to detect PSTV rapidly.The results indicated that when the anneal temperature was 37℃ and the circles were more than 10 times in reverse-transcription phase,a relative band of PSTV was obtained by PCR which could be proved using nested-set PCR.Single-enzyme RT-PCR was simple and low-cost,so it could play on important role for the detection of PSTV.
Key words:
PSTV,
Single-enzyme RT-PCR,
Taqenzyme
中图分类号:
叶明, 盛长忠, 石明, 安春菊, 冀俊丽, 吴学锋, 杜荣骞, 李德森. 单酶法RTP-CR检测马铃薯纺锤块茎类病毒[J]. 华北农学报, 2002, 17(1): 41-45. doi: 10.3321/j.issn:1000-7091.2002.01.009.
Ye Ming, Sheng Changzhong, Shi Ming, An Chunju, Ji Junli, Wu Xuefeng, Du Rongqian, Li Desen. Detection of Potato Spindl a Tuber Viroid by Single- enzyme RT-PCR[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2002, 17(1): 41-45. doi: 10.3321/j.issn:1000-7091.2002.01.009.