CAJ | 학술논문

用绵羊睾丸细胞培养山羊痘病毒疫苗株G14-STV44-55,提取其基因组为模板,设计TK基因的特异性引物并进行PCR扩增,获得了2 405 bp的DNA片段,并将PCR产物克隆至pGEM-T Easy载体.酶切鉴定、PCR鉴定和测序结果表明,成功获得了TK基因,获得的TK基因内存在唯一的ACC65I酶切位点和3′末端早期转录终止信号TTTTTN(T)T.核苷酸和氨基酸同源性分析表明,弱毒疫苗株G14-STV44-55TK基因与基因Bank中发表的13个参考毒株的同源性在96%和95%以上,说明羊痘病毒TK基因具有高度的保守性.
용면양고환세포배양산양두병독역묘주G14-STV44-55,제취기기인조위모판,설계TK기인적특이성인물병진행PCR확증,획득료2 405 bp적DNA편단,병장PCR산물극륭지pGEM-T Easy재체.매절감정、PCR감정화측서결과표명,성공획득료TK기인,획득적TK기인내존재유일적ACC65I매절위점화3′말단조기전록종지신호TTTTTN(T)T.핵감산화안기산동원성분석표명,약독역묘주G14-STV44-55TK기인여기인Bank중발표적13개삼고독주적동원성재96%화95%이상,설명양두병독TK기인구유고도적보수성.
The capripoxvirus live attenuated vaccine strain G14-STV44-55 was cultured with sheep testicular cell, and the genomic DNA was extracted from the virus strain, and a pair of specific primers were desined in order to amplify a TK gene. The PCR product approximately 2 405 bp in size was cloned into pGEM-T Easy vector . Restriction enzyme assay ,PCR and sequencing confirmed that the TK gene was obtained successfully. Only one ACC65 I site and a transcription termination signal TTTTTN ( T) T were found in 3′ side of TK gene. Analysis showed that G14-STV44-55 strain shared 96 % and 95 % identity rates with the reference strains in levels of nucleotide and amino acid .It indicate that TK gene is very conservative.