CAJ | 학술논문

根据从虎气管组织扩增获得的猫传染性鼻气管炎病毒相关核酸序列，设计扩增编码虎源猫传染性鼻气管炎病毒 gD 蛋白基因片段的引物，通过 PCR 扩增出 gD 基因片段，并成功插入到 pMD18-T Sim-ple 载体中，筛选获得重组质粒，命名为18T-gD。重组质粒通过 Hin d Ⅲ和 Xho Ⅰ酶切后，插入 pcDNA 3．1（＋）真核表达载体构建出 gD 基因的重组真核表达质粒 pcDNA-gD。用脂质体将重组真核表达质粒 pcD-NA-gD 转染293T 细胞，通过 RT-PCR 和 Western blot 检测，结果显示 gD 基因在真核细胞中得到正确转录和表达，所表达蛋白分子质量约为42．29 ku，为虎源猫传染性鼻气管炎基因疫苗的研究奠定了基础。
근거종호기관조직확증획득적묘전염성비기관염병독상관핵산서렬，설계확증편마호원묘전염성비기관염병독 gD 단백기인편단적인물，통과 PCR 확증출 gD 기인편단，병성공삽입도 pMD18-T Sim-ple 재체중，사선획득중조질립，명명위18T-gD。중조질립통과 Hin d Ⅲ화 Xho Ⅰ매절후，삽입 pcDNA 3．1（＋）진핵표체재체구건출 gD 기인적중조진핵표체질립 pcDNA-gD。용지질체장중조진핵표체질립 pcD-NA-gD 전염293T 세포，통과 RT-PCR 화 Western blot 검측，결과현시 gD 기인재진핵세포중득도정학전록화표체，소표체단백분자질량약위42．29 ku，위호원묘전염성비기관염기인역묘적연구전정료기출。
The primers of gD genes of the FHV-1 were designed according to sequences of the FHV-1 from GenBank,gD gene was amplified by PCR and cloned successfully into pMD18-T Simple vector.gD gene was constructed into eukaryotic expression vector pcDNA 3.1(+),the recombinant plasmid ,named as pcDNA-gD,was transfected into 293T cells.The results of RT-PCR and Western blot indicated that the gD gene was transcripted and expressed,and the recombinant fusion protein was about 42.29 ku.The results lay an important foundation for the further research of FHV-1.