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Basic research for mini-pig production to induction og human MHC genes

小型猪生产诱导人类MHC基因的基础研究

基本信息

批准号：
09470260
负责人：
SADA Masaharu
金额：
$ 6.59万
依托单位：
National Cardiovascular Center
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470260/
关键词：
CSK minipig CLAWN minipig HLA class II genes SLA-DR genes SLA-DQ genes gene cloning SBT xeno-transplanation SLA遺伝子 SLA-DMA遺伝子 HLA-DQB遺伝子 SLA-DQB遺伝子 PCR-LiPA genotyping CSK系ミニブタ NIHミニブタ

项目摘要

The swine major histocompatibility complex (SLA) is encoded a large number of genes including the class I, II, and III on swine chromosome 7. Structural and functional properties of the SLA confirm the similarity to that of human major histocompatibility complex (HLA). In this study, we have isolated a full-length cDNA clone of SLA class II (DRB, DQB, DQA, DNA) genes of CSK minipig (modified Gottingen) and CLAWN minipig (originated Kagoshima Univ.) , and analyzed the coincidence of amino acid sequence between SLA class II genes and HLA class II genes to induction of HLA genes to minipig.The mRNA was extracted from CSK and CLAWN minipig spleen. The cDNA library was constructed and screened by the HLA class II cDNA clone as a probes. These clone encoded a primary translated products of 250 amino acids, which includes a signal peptide of 27 amino acids were compared to amino acid sequence of HLA class II genes.Amino acid comparative analysis between SLA class II genes and HLA class II genes were exhibited a high degree of overall identify to the HLA-DRB, -DRA, -DQA, and -DNA molecules (75%, 87%, 86%, 83% identities, respectively). The analysis of SLA genes is important to develop a minipig to induction of HLA genes for conducting forthcoming xenotransplant donor.

猪主要组织相容性复合体（SLA）在猪7号染色体上编码了大量的基因，包括I、II和III类基因。SLA的结构和功能特性与人类主要组织相容性复合体（HLA）相似。本研究分离了改良哥丁根的CSK迷你猪和鹿儿岛大学的CLAWN迷你猪的SLAⅱ类（DRB、DQB、DQA、DNA）基因的全长cDNA克隆，分析了SLAⅱ类基因与HLAⅱ类基因在诱导迷你猪HLA基因中的氨基酸序列一致性。从小型猪CSK和CLAWN脾脏中提取mRNA。构建cDNA文库，以HLAⅱ类cDNA克隆为探针进行筛选。这些克隆编码了250个氨基酸的初级翻译产物，其中包括27个氨基酸的信号肽，并与HLA II类基因的氨基酸序列进行了比较。SLAⅱ类基因与HLAⅱ类基因的氨基酸比较分析显示，对HLA- drb、-DRA、-DQA和-DNA分子具有高度的总体识别度（分别为75%、87%、86%和83%）。SLA基因的分析对于培育小型猪来诱导HLA基因进行即将到来的异种移植供体具有重要意义。