湖南沙子岭猪内源性逆转录病毒的研究

遗传 ›› 2006, Vol. 28 ›› Issue (7): 799-804.

湖南沙子岭猪内源性逆转录病毒的研究

邢晓为1，薛立群2，黄生强2，黎淑娟1，王维1

1. 中南大学湘雅三医院细胞移植与基因治疗研究所，长沙 410013；
2. 湖南农业大学动物科学技术学院，长沙 410128

收稿日期:2005-09-19 修回日期:2005-11-09 出版日期:2006-07-10 发布日期:2006-07-10
通讯作者: 王维

Investigation of the biological characters of porcine endogenous retrovirus (PERV) in Shazi Ling pigs

XING Xiao-Wei 1, XUE Li-Qun 2, HUANG Sheng-Qiang 2, LI Shu-Juan1,WANG Wei1

1. Cell Transplantation & Gene Therapy Center, The Third Xiangya Hospital of Central South University, Changsha 410013,China; 2. Department of Animal Science and Technology, Hunan Agriculture University, Changsha 410128, China

Received:2005-09-19 Revised:2005-11-09 Online:2006-07-10 Published:2006-07-10
Contact: WANG Wei

摘要/Abstract

摘要：

为评价从猪到人异种移植的生物安全性提供依据，从湖南沙子岭猪的保种群内随机采集31头个体的耳样组织，应用PCR和RT-PCR技术分别检测这些组织中内源性逆转录病毒（porcine endogenous retrovirus，PERV）的前病毒DNA和mRNA，并对PCR扩增的灵敏性进行评估。多组织RT-PCR检测3头沙子岭猪肾、心、肝、肺、脾等组织中PERV的表达情况，了解其在各组织中的分布情况；最后，扩增、测序该猪种的env基因，结果用NCBI中的BLAST软件进行分析。PCR和RT-PCR结果表明，所检测的31头沙子岭猪均带有PERV前病毒DNA，耳样组织中均有PERV mRNA表达，其中有2头个体携带 env-A、env-B、env-C 3种囊膜蛋白基因，而其余的29头个体只带有env-A、env-B 两种囊膜蛋白基因，未检测到env-C基因。多组织RT-PCR扩增结果表明，3头沙子岭猪的肾、心、肝、肺、脾等组织中，pol、gag、env-A、env-B 基因均有表达，未检测到env-C基因表达。测序沙子岭猪的env基因，结果发现，沙子岭猪env-B 和env-C基因与其他猪种序列比较分别存在2 和10个碱基的差异，而env-A基因序列没有差异，说明不同的猪种之间 env基因存在多态性。以上结果表明，沙子岭猪种群携带PERV，其亚型主要以PERV-A，B为主；PERV在该猪种肾、心、肝、肺、脾等多种组织中的分布没有明显组织特异性，且93.5 % (29/31)个体表现为 env-C 基因缺失，提示沙子岭猪作为候选猪种可能在异种移植中具有较好的应用前景。

关键词: 猪内源性逆转录病毒(PERV), 沙子岭猪, 亚型, 异种移植

Abstract:

To provide basic parameters of evaluating the biological safety of xenotransplantation from pig to human, ear tissues from 31 individuals were randomly collected from a Shazi Ling pig population. PCR and RT-PCR were performed to detect porcine endogenous retrovirus (PERV) proviral DNA and mRNA respectively. The sensitivity of the PCR was evaluated using a positive control. To study tissue distribution, RT-PCR of pol, gag and env was performed in the kidney, heart, liver, lung and spleen of 3 individuals. Finally, env-A, env-B and env-C were amplified, sequenced and analyzed using the BLAST software in NCBI (National Center for Biotechnology Information). The results showed PERV proviral DNA and mRNA could be detected in all 31 individuals by PCR and RT-PCR, respectively. env-A, env-B and env-C were only detected in 2 individuals, while in the other 29 individuals, only env-A and env-B but not env-C was detected. The quantity of DNA in PCR amplification of PERV genes should be more than 15 ng. RT-PCR results showed gag, pol, env-A and env-B were expressed in the kidney, heart, liver, lung and spleen of all 3 individuals, but env-C was not. Sequencing of env genes in Shazi Ling pigs revealed that while there was no difference in env-A sequence when compared to other herd in GenBank, there were 2 and 10 bp differences in the sequences of env-B and env-C respectively, suggesting that env gene is polymorphic in different pig strains. PERV exists in the Shazi Ling pig population and the predominant subtype is PERV-A, B. The distribution of PERV displays no significant tissue specificity and env-C is absent in 93.5 % (29/31) of the individuals. The results indicate that Shazi Ling pig may have great potential value as a candidate in xenotransplantation.

中图分类号:

邢晓为，薛立群，黄生强，黎淑娟，王维. 湖南沙子岭猪内源性逆转录病毒的研究 [J]. 遗传, 2006, 28(7): 799-804.

XING Xiao-Wei , XUE Li-Qun , HUANG Sheng-Qiang , LI Shu-Juan, WANG Wei . Investigation of the biological characters of porcine endogenous retrovirus (PERV) in Shazi Ling pigs[J]. HEREDITAS, 2006, 28(7): 799-804.

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