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反义RNA对猪α-1,3-半乳糖苷转移酶活性的影响

The Effect of Antisense RNA on Activity of Porcine α-1,3-Galactosyltransferase

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【作者】 林丛许克锋张文秀侯宽永毛泽斌

【Author】 LIN Cong 1) , XU Ke feng 1) , ZHANG Wen xiu 1) , HOU Kuan yong 1) , MAO Ze bin 2)* ( 1) Department of General Surgery, Peking University Third Hospital, Beijing 100083, China; 2) Department of Biochemistry and Molecular

【机构】 北京大学第三医院普外科!北京100083北京大学医学部生物化学与分子生物学系!北京100083

【摘要】 α 1,3 半乳糖表位是猪 人异种移植超急性排斥反应的主要抗原 ,由α 1,3 半乳糖苷转移酶催化合成 .用RT PCR方法扩增中国实验用小型猪α 1,3 半乳糖苷转移酶cDNA的前 582bp ,测定碱基序列并构建其反义表达载体pLXRN ,将其转染入猪主动脉内皮细胞 .NorthernBlotting表明α 1,3 半乳糖苷转移酶mRNA减少 .检测α 1,3 半乳糖苷转移酶活性表明 ,反义RNA可使其活性下降32 2 % .研究结果表明可能通过反义RNA来抑制猪 人异种移植超急性排斥反应

【Abstract】 In pig to primate xenotransplantation, pig organs were rejected hyperalcutely by antibody mediated complement activation. α 1,3 galactose (αGal) epitopes on pig vascular endothelium were the major targets of hyperacute rejection. This epitope was produced by α 1,3 galactosyltransferase(α1,3GT). The first 582bp cDNA of α 1,3 galactosyltransferase was amplified by RT PCR, sequenced and cloned into retroviral expression vector pLXRN in antisense direction. After in vitro transfection into porcine aortic endothelial cells(PAECs), it was shown that the α 1,3 galactosyltransferase mRNA in PAECs decreased and the activity of α 1,3 galactosyltransferase was reduced by 32.2%. So antisense RNA may be a possible way of inhibiting hyperacute rejection in pig to primate xenotransplantation.

【基金】 国家自然科学基金 (No .30 0 0 0 199)资助&&
  • 【文献出处】 中国生物化学与分子生物学报 ,Chinese Journal of Biochemistry and Molecular Biology , 编辑部邮箱 ,2001年04期
  • 【分类号】Q555;R617
  • 【被引频次】4
  • 【下载频次】79
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