摘要
随着现代分子生物学技术的发展,反转录聚合酶链式反应技术在甘薯病毒检测上的应用越来越广泛。采用该技术可检测出甘薯组织中含量极低的病毒RNA,具有灵敏度高、特异性强等优点。在本研究中根据NCBI Gen Bank中收录的SPCSV病毒外壳蛋白(CP)基因核苷酸序列的保守区域设计了3对特异性引物,使用天根生化科技(北京)有限公司生产的Quant One Step RT-PCR kit试剂盒,针对影响扩增产物的影响因素退火温度进行优化,建立了甘薯退绿矮化病毒的RT-PCR检测方法。该方法使RT-PCR在一管内完成,大大降低了外源物质的污染及RNA降解的几率,可作为甘薯SPCSV病毒的快速检测方法。
With the development of modern molecular biological technology, reverse transcription-polymerase chain reaction has been widely used in detection of sweet potato viruses. Little RNA viruses from sweet potato tissue can be detected by RT-PCR which are with such advantage as more sensitive and specificity. The primers were designed and synthesized according to the published nucleotide sequence of coat protein gene of sweet potato chlorotic stunt virus（ SPCSV）, and the detection procedure of reverse transcription-polymerase chain reaction（RT-PCR） for SPCSV was built in this paper based on the optimization of the annealing temperature which influxes amplified products with the products made by Tiangen Biotech（Beijing） Co. LTD. The method can make reverse transcription-polymerase chain reaction finished in one tube, and significantly reduces the effect of exogenous substances contamination and the risk of RNA degradation, so this way can quickly finish the detection of sweet potato chlorotic stunt virus.
作者
肖海峻
孟利前
陈梦然
杨春林
Xiao Haijun Meng Liqian Chen Mengran Yang Chunlin(Beijing Vocational College of Agriculture, Beijing, 102442)
出处
《分子植物育种》
CAS
CSCD
北大核心
2017年第7期2896-2900,共5页
Molecular Plant Breeding
基金
北京农业职业学院院级课题资助
