登革病毒(NGC株)C与prM全长基因的扩增与克隆  

Amplification and cloning the full-length C gene and prM gene from dengue virus(NGC strain)

作　　者：葛春喜[1] 陈观今[1] 黄炯烈[1] 

机构地区：[1]中山医科大学病原生物学部,广州510080

出　　处：《中国人兽共患病杂志》2002年第1期5-7,共3页Chinese Journal of Zoonoses

摘　　要：目的　从登革病毒 (NGC株 )中快速分离基因组RNA ,RT -PCR扩增及克隆C和 prM全长基因 ,从而为研究蚊虫细胞对登革病毒的细胞内免疫研究奠定基础。方法　以Trizol试剂从C6 / 36细胞培养上清中提取基因组RNA ,RT -PCR扩增全长C和 prM基因 ,T -A重组入pGEM -T载体 ,重组质粒的双酶切、PCR与测序鉴定。 结果与结论　应用Trizol试剂从蚊细胞培养液上清中快速分离到高纯度和完整登革病毒RNA基因组 ,成功扩增与克隆出全长登革病毒C和Aim To extract the dengue virus RNA genome rapidly from liquid medium for mosquito culture cell C6/36 and amplify and clone the full length C and prM gene.Methods Extract the dengue virus RNA genome by using the Trizol reagent.Two pairs of primers were designed according to the DV 2 sequences.By using RT-PCR technique,C and prM gene were amplified and linked to pGEM T vector.Then they were transformed to E.coli JM109.The recombinant plasmid was identified by PCR and Sac I and Kpn I digestion.The recombinant plasmid was also identified by sequencing.Result and Clonclusions The high purified and intact virus RNA genome was extracted by Trizol reagent.The full length C and prM gene were amplified and cloned.

关 键 词：登革病毒 NGC株 prM基因 C基因 RT-PCR 

分 类 号：R373.33[医药卫生—病原生物学]