中国两头乌猪品种内源性逆转录病毒基因研究

目的对5个中国两头乌猪品种(通城猪、东山猪、沙子岭猪、赣西两头乌猪和金华猪)及3个国外品种(大白猪、长白猪和杜洛克猪)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)基因的3个亚型A、B、C,分别从DNA和RNA水平上进行研究,以发现中国两头乌猪品种在异种器官移植中的资源优势。方法利用PCR方法在DNA水平上对PERV基因的三个亚型进行鉴定,并通过半定量PCR方法在RNA水平上检测通城猪和大白猪PERV各亚型在心、肝、脾、肺、肾、肌肉、脂肪、淋巴和脑组织中的表达谱。结果4个华中两头乌猪种中env-AB型为主要PERV亚型,分别占被测总数的92%～100%。在这4个品种中均没有检测到C亚型,金华猪以及3个国外猪种中均检测到了C亚型,病毒亚型种类也更丰富。半定量PCR实验结果显示gag、pol基因在两个品种9个组织中广泛表达,env-A在通城猪的心、肝、肺、脂肪和淋巴组织中表达量较低,env-B在通城猪的心脏和淋巴组织中表达量较低,而env-B在大白猪的肾脏中表达很低,其他所测8个组织中表达量都较高。结论通城猪、东山猪、赣西两头乌猪和沙子岭猪可以做为较佳的异种移植候选供体,具有良好的应用前景。

Identification of porcine endogenous retrovirus in Chinese two-end-black pigs

Objective In order to explore whether the Chinese two-end-black pigs are superior for xenotransplanration, the gag, pol and env genes of porcine endogenous retrovirus （PERV） in five Chinese two-end-black pig breeds （Tongcheng pig, Dongshan pig, Ganxi two-end-black pig, Shaziling pig and Jinhua pig） and three Western breeds （Yorkshire, Landrace and Duroc） were identified at DNA and RNA levels. Methods The presence of gag, pol and env-A, B, and C genes of PERV was detected by PCR, and the expression spectrum of gag, pol and env gcne of PERV in the heart, liver, spleen, lung, kidney, muscle, adipose, lymphatic and brain tissues of Tongcheng and Yorkshire pigs was determined by semi-quantitative RT-PCR. Results In four breeds of two-end-black pigs in Central China, the major subtype of PERV was env-AB, with a positive rate of 92% to 100%. Gene subtype Env-C was not identified in all these four breeds, but was found in the Jinghua pig and three Western breeds. The results of semi-quantitative PCR showed that Gag and pol genes were widely expressed in all the nine tissues. In the Tongcheng pigs, env-A expression in the heart, liver, lungs, adipose and lymphatic tissues was lower than that in the other tissues, while env-B expression was very low in the kidneys hut higher in the other 8 tissues of Yorkshire pigs. Conclusions The results of our study indicate that four Chinese two-endblack pig breeds （Tongcheng pig, Dongshan pig, Ganxi two-end-black pig, Shaziling pig） may become good donor candidate for xenotransplantation, and have promising future of application.