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长白与青海八眉“二元杂交”猪种群PERV亚型分析
  • ISSN号:1002-2694
  • 期刊名称:《中国人兽共患病学报》
  • 时间:0
  • 分类:R617[医药卫生—临床医学;医药卫生—外科学]
  • 作者机构:[1]中南大学湘雅三医院细胞移植与基因治疗中心,长沙410013, [2]青海大学农牧学院
  • 相关基金:国家自然基金项目(30371493),湖南省卫生厅项目(132005067)
中文摘要:

目的研究长白与青海八眉“二元杂交”猪种群携带的内源性逆转录病毒(porcine endogenous retrovirus,PERV)亚型,为评价从猪到人异种移植的生物安全性提供依据。方法从长白与青海八眉“二元杂交”猪种群中随机采集33头个体外周血,应用PCR和RT-PCR技术分别检测PERV前病毒DNA和mRNA,研究该种群携带PERV的亚型,并对PCR扩增的灵敏性进行评估。克隆测序该猪种PERVenv基因,结果用NCBI中的BLAST软件进行分析。结果所检测的33头个体均带有PERV前病毒DNA,外周血中均有PERV mRNA表达,其中,69.7%(23/33)个体携带env-A,env-B和env-C三种囊膜蛋白基因,而其余的30.3%(10/33)个体只带有env-A和env-B两种囊膜蛋白基因,env-C基因缺失。灵敏性分析实验结果表明,PCR扩增PERV时,DNA模板不能少于15ng。克隆env-A,env-B和env-C基因,结果提交GenBank(GenBank登录号分别为DQ856326、DQ856327、DQ856328)。生物信息分析结果表明,该猪种env基因与GenBank登录其他猪种序列相比存在差异,其中env-A、env-B基因发生移码突变,蛋白合成提前终止。结论长白与青海八眉“二元杂交”猪种群携带PERV,69.7%个体为PERV-ABC亚型;缺失突变使PERVenv-A,env-B基因阅读框发生改变,有可能影响其感染特性,从生物安全性方面综合考虑,该猪种不宜作为异种移植供体。

英文摘要:

To investigate the subtypes of porcine endogenous retrovirus (PERV) in hybrids of Landrace with Qinghai Bamei swine, so as to provide basic parameters to evaluate the biological safety of xenotransplantation of pig to human, the peripheral blood samples from 33 individuals were randomly collected from hybrids of Landrace with Qinghai Bamei swine, and PCR and RT-PCR were used to detect the provital DNA and mRNA of PERV respectively. The sensitivity of the PCR assay was evaluated using a positive control, and the PERV env gene was amplified, sequenced and analysed by using the BLAST software in National Center of Biotechnology Information (NCBI). The results showed that the provital DNA and mRNA of PERV could be detected in all these 33 individuals by means of PCR and RT-PCR, in which 69.7% (23/33) of individuals tested carried the envelop protein genes envA, envB and envC , while the other 30.3% of individuals carried only envA and envB, but no envC. It was demonstrated that the quantity of DNA to be detected by PCR amplification of the PERV genes was not less than 15 ng. These PERV envA, envB and envC genes were sequenced and submitted to GenBank with GenBank accession numbers of DQ856326, DQ856327, DQ856328 respectively. The information analysis showed that there were significant differences in env genes comparing with other breeds in GenBank, in which frameshift mutation occurred in envA and envB genes, thus causing premature stop of protein synthesis. It is concluded that from the aspect of biological safety, this breed is not suitable as donor in xenotransplantation, because 69.7% of individuals are the subtypes of PERV-A, B,C, in which frameshift mutation may occur, thus changing the character of the cross-species transmmision of PERV.

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期刊信息
  • 《中国人兽共患病学报》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中国微生物学会
  • 主编:严延生
  • 地址:福州市津泰路76号
  • 邮编:350001
  • 邮箱:rsghb@fjcdc.com.cn
  • 电话:0591-87552018
  • 国际标准刊号:ISSN:1002-2694
  • 国内统一刊号:ISSN:35-1284/R
  • 邮发代号:34-46
  • 获奖情况:
  • 中国基础医学核心期刊,中国生物医学核心期刊,中国科学引文数据库列为来源期刊及统计源,中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),英国农业与生物科学研究中心文摘,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:9000