摘要
用自行设计的 1对引物 you1+和 you2 -对 5个传染性支气管炎病毒 (IBV)地方分离株 (HN2 ,HN4,JX1,SC2和SC4)的基因组 RNA进行 RT- PCR,均成功地扩增出预期大小 (约 16 30 bp)的目的片段 ,经 PCR、酶切鉴定为 S1基因。将 5个毒株的 S1基因克隆到 p GEM- T载体中 ,重组质粒经 Eco R 和 Hind 单酶切以及用引物 you1+和 you2 -进行质粒 PCR扩增鉴定 ,证明 IBV- S1基因已成功地克隆到了 p GEM- T载体中。同时 ,为了测序的方便 ,还将 IBV- S1基因作了目的片段 (约 10 6 0 bp)
RT-PCR was used to amplify the S1 gene fragments of the five Chinese isolates of IBV(HN2,HN4,JX1,SC2 and SC4) using the primers you1+ and you2-.The result indicated the size of the RT-PCR products was the same as expected(about 1.6 kb).The RT-PCR products were used as templet to amplify with primers you1+ and you2- and digested with the restriction enzyme PstⅠ.All the results demonstrate that the S1 gene fragments of the five IBV isolates have been attained successfully.The S1 genes of five isolates were cloned to pGEM-T vector,and then its subclones were made successfully so as to sequence more conviniently.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2002年第3期222-225,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金重大项目 ( 398932 90 -2 B)
