CAJ | 학술논문

为了建立检测猪内源性反转录病毒(porcine endogenous retrovirus,PERV)的特异性方法,根据已发表的PERV的序列,设计并合成了针对PERV核心蛋白(gag)、多聚酶(pol)、囊膜蛋白(env)基因的3对引物,预期扩增片段分别为361、150、265 bp.应用PCR技术检测了PERV在4株猪源细胞中的整合情况.结果表明,在所有被检细胞的基因组中均存在有PERV的前病毒序列;应用RT-PCR检测上述4株猪源细胞中PERV特异性mRNA的表达,结果均为阳性.试验还对建立的上述2种方法的特异性进行了探讨,结果表明试验建立的PERV检测法具有较高的特异性.该方法的建立为进一步研究PERV奠定了基础,还可对异种移植动物模型及异种移植受体进行病原安全性监测.
위료건립검측저내원성반전록병독(porcine endogenous retrovirus,PERV)적특이성방법,근거이발표적PERV적서렬,설계병합성료침대PERV핵심단백(gag)、다취매(pol)、낭막단백(env)기인적3대인물,예기확증편단분별위361、150、265 bp.응용PCR기술검측료PERV재4주저원세포중적정합정황.결과표명,재소유피검세포적기인조중균존재유PERV적전병독서렬;응용RT-PCR검측상술4주저원세포중PERV특이성mRNA적표체,결과균위양성.시험환대건립적상술2충방법적특이성진행료탐토,결과표명시험건립적PERV검측법구유교고적특이성.해방법적건립위진일보연구PERV전정료기출,환가대이충이식동물모형급이충이식수체진행병원안전성감측.
Porcine endogenous retroviruses (PERV) can infect human cell in vitro, which raised widely concerns re-garding the transmission of PERV to xenograft recipients. It's essential to establish a method for detection of PERV.3 pairs of primers were synthesized according to the sequence of gag, pol and env gene of PERV. Polymerase chainreaction (PCR) and reverse transcription PCR (RT-PCR) assays were performed for detection of PERV provirusDNA and PERV specific mRNA. The results showed that provirus DNA and mRNA of PERV existed and expressedin all 4 tested cell lines. The sizes of amplified fragments are identical with the predicted. These methods may be suit-able for monitoring PERV in other cells or tissue.