梨树苹果茎痘病毒PRINS检测中引物的设计及退火温度的测定  

The Design of the Primer and the Testing of Annealing Temperature of ASPV PRINS Detection

作　　者：吴庆丰[1] 牛建新[1] 李文慧[1] 刘娜[1] 

机构地区：[1]石河子大学农学院,新疆石河子832003

出　　处：《新疆农业科学》2011年第11期2067-2070,共4页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金项目(30360066);国家科技攻关计划引导项目(2003BA546C);石河子大学自然科学与技术创新(500002125707)

摘　　要：【目的】利用5条引物目的片段进行扩增,完成PRINS检测中引物及其退火温度的测定。【方法】采用确认带有ASPV的库尔勒香梨叶片,以叶片为试验材料,并采用经RT-PCR检测技术健康香梨的叶片作为阴性对照。引物采用NCBI提供的BLAST工具中的ASPV病毒的全基因组序列的外壳蛋白的保守区域进行设计。并根据PRINS特有的性质及ASPV病毒有片断相互重叠现象来设计相应的引物。【结果】这5条引物之间没有太多的干扰,且都在目的区域出现了比较清晰的条带。【结论】证明所设计引物可以在进一步的PRINS检测ASPV病毒时直接使用,为PRINS在梨树病毒检测方面发挥更大的作用奠定了基础。[ Objective ] In order to complete testing PRINS primer and annealing temperature measurement, five primers were used to amplify the purpose fragments separately. [ Method ] Korla Pear leaves identified with ASPV were selected as test materials, healthy pear leaves detected by RT -PCR were selected as negative control. Primers were designed by using the conserved regions of the coat protein of ASPV virus＇ complete genome sequence, which was part of the BLAST tool provided by NCBI. In addition, the distinctive characteristics of PRINS and the phenomenon that ASPV fragments overlapped with each other were also taken into consideration in the process of primer design. [ Result ] There was little influence between five primers. And clear bands were all showed on the purpose locations. [ Conclusion ] Therefore these five primers could be used directly for the next step of PRINS detection. It laid a good foundation for PRINS to detect pear virus.

关 键 词：引物原位标记 引物 病毒 检测 

分 类 号：S661.2[农业科学—果树学] S188.2[农业科学—园艺学]