摘要
参考Genbank收录的IBV纤突蛋白 (S1)基因序列 ,自行设计合成一对引物 ,对传染性支气管炎病毒 (IBV)江苏省地方分离毒株 (JS/95/0 3)RNA进行RT PCR扩增 ,产物经琼脂糖凝胶电泳分析 ,呈现一条 1716bp的条带 ,将其克隆入T/A质粒pMD18 T载体中 ,转化大肠杆菌JM10 9,挑选阳性克隆 ,用质粒少量提取法提取重组质粒 ,用EcoRⅠ和HindⅢ双酶切对重组克隆质粒进行鉴定 ,然后进行序列测定 ,证实为S1基因。将此重组质粒命名为pMDJS950 3S。
A pair of primers were designed and synthesized according to the reported spike protein(S1)gene sequence of IBV.Then the viral RNA of IBV(JS/95/30)was amplified by reverse transcription polymerase chain reaction(RT PCR).The amplified products were analyzed by agarose gel electrophoresis,and all appeared a fragment of 1 716 bp as expected.The RT PCR products were cloned into the T/A plasmid pMD18 T.The recombinant plasmid was digested with restriction endonuclease EcoRⅠ and HindⅢ,and the recombinant plasmid was also sequenced.The result suggested that it be the S1 gene of IBV.We designated the recombinant plasmid as pMDJS9503S.
出处
《畜牧与兽医》
北大核心
2000年第5期1-3,共3页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金重大项目 !(398932 90 2 2 )资助
