摘要
根据猪内源性逆转录病毒(PERV)的核心蛋白(gag)基因、多聚酶(pol)基因、囊膜(env)基因和β-肌动素(β-actin)基因核酸序列设计并筛选特异性引物6对,利用PCR方法对五指山小型猪PERVgag、pol和env基因存在情况进行系统检测;利用半定量RT-PCR方法对五指山小型猪PERVgag、pol和env基因在心、肝、脾、肺、肾、脑、肌肉7个组织中的表达情况进行系统检测与分析。结果显示,在202个被检样品中,gag、pol基因和envB基因亚型均存在,envA基因亚型仅3个样品未发现,envC基因亚型共检出34个,检出率为16.8%。所测试的7个组织均表达gag、pol和env基因序列,但envC仅在1头个体的组织中表达。各组织gag mRNA丰度以肾脏为最高,其次为肺、心、肝、脾,脑和肌肉组织的丰度最低。各组织pol mRNA丰度以肾脏为最高,其次为脾、肺、肝、心,肌肉和脑组织的丰度最低。各组织envA、envB mRNA丰度信号强度分布与gag、pol信号类似,亦以肾脏为最高,脑和肌肉组织的丰度最低。各组织envC mRNA的信号强度明显低于envA、envB。以上结果表明,在五指山小型猪中存在PERV,且在各组织中的表达量有差异。
According to the nucleotide sequences of gag,pol and env gene of PERV and β-actin gene,six pairs of specific primer were desired and synthesized. It was detected systemically the existence of gag, pol and env gene of PERV in the Wuzhis-han miniature pig by PCR and the expression level of gag, pol and env gene of PERV in seven tissues by semi-quantitative RT-PCR.The results showed that gag,pol and envB gene subtype existed in all 202 samples,envA gene subtype didn＇ t exist only in three samples,thirty-four samples were detected for envC gene subtype with the positive rate of 16.8% ;gag, pol, env mRNA expressed in these seven tissues,but envC mRNA expressed only in one pig.The signal of gag mRNA was the highest was in kidney, the lower in lung,heart,liver,spleen,the lowest in brain and muscle.The signal of pol mRNA was the highest also in kidney,the lower in spleen,lung,liver,heurt, the lowest in brain.The signals of envA and envB mRNA were similar with that of gag and pol, the highest was in kidney,the lowest in brain and muscle. The signal of envC mRNA was distinctly lower than that of envA and en-vB,indicating that PERV do exist and expresse differently in each tissue in the Wuzhishan miniature pig,This survey may be of great value for Wuzhishan miniature pigs being used for the xenotransplantation and assessing the pathogen safety of PERV.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2008年第1期68-71,93,共5页
Chinese Journal of Veterinary Science
基金
国家“863”高科技研究发展计划资助项目(2003AA205104)
