Chungnam National University, Daejeon, Korea.
Anim Genet. 2010 Feb;41(1):89-92. doi: 10.1111/j.1365-2052.2009.01953.x. Epub 2009 Sep 24.
Xenotransplantation from pigs provides a possible solution to the shortage of human organs for allotransplantation. Porcine endogenous retroviruses (PERVs) are a possible obstacle to using porcine organs in addition to the immunological barriers. Three main types of PERVs (A, B and C) have been previously investigated in diverse pig breeds. To examine the copy numbers of PERVs and their genomic locations in the Korean native pig genome, we screened a BAC (Bacterial Artificial Chromosome) library with PERV-specific protease primers for initial recognition of PERV-positive clones and three sets of envelope-specific primers for the identification of PERV types. A total of 45 PERV-positive clones, nine PERV-A and 36 PERV-B, have been identified from the library screening and the BAC contigs were constructed using the primers designed from BAC end sequences (BESs). These primers were also used for SCH (Somatic Cell Hybrid) and RH (Radiation Hybrid) mapping of the PERV-positive clones. The results indicate that 45 PERV-positive BAC clones belong to nine contigs and a singleton. SCH and IMpRH (INRA-Minnesota Porcine Radiation Hybrid) mapping results indicated that there are at least eight separate PERV genomic locations, consisting of three PERV-A and five PERV-B. One contig could not be mapped, and two contigs are closely located on SSC7. Southern blotting indicates there may be up to 15 additional sites. Further investigation of these clones will contribute to a general strategy to generate PERV-free lines of pigs suitable for xenotransplantation.
异种移植为同种异体移植提供了一种可能的解决方案,以解决人类器官短缺的问题。除了免疫屏障外,猪内源性逆转录病毒(PERVs)也是使用猪器官的一个可能障碍。以前已经在不同的猪品种中研究了三种主要类型的 PERV(A、B 和 C)。为了研究 PERV 在韩国本土猪基因组中的拷贝数及其基因组位置,我们用 PERV 特异性蛋白酶引物筛选了 BAC(细菌人工染色体)文库,用于初步识别 PERV 阳性克隆,并用三组包膜特异性引物识别 PERV 类型。从文库筛选中总共鉴定出 45 个 PERV 阳性克隆,其中 9 个为 PERV-A,36 个为 PERV-B,并用从 BAC 末端序列(BES)设计的引物构建了 BAC 连续体。这些引物也用于 PERV 阳性克隆的 SCH(体细胞杂交)和 RH(辐射杂交)作图。结果表明,45 个 PERV 阳性 BAC 克隆属于九个连续体和一个单一体。SCH 和 IMpRH(INRA-Minnesota 猪辐射杂交)作图结果表明,至少有八个独立的 PERV 基因组位置,包括三个 PERV-A 和五个 PERV-B。一个连续体无法作图,两个连续体在 SSC7 上紧密相邻。Southern 印迹表明可能还有多达 15 个其他位点。对这些克隆的进一步研究将有助于制定生成适合异种移植的无 PERV 猪系的一般策略。