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机构地区:[1]四川农业大学动物生物技术中心,四川雅安625014
出 处:《黑龙江畜牧兽医》2007年第5期18-20,共3页Heilongjiang Animal Science And veterinary Medicine
基 金:国家"973"项目(2004CCA01800)
摘 要:根据GenBank收录的猪白细胞介素-6(PIL-6)设计1对特异性引物,经刀豆蛋白素A(Co—nA)诱导猪淋巴细胞并提取总RNA,用1it—PCR方法扩增出荣昌猪IL-6的cDNA。将扩增基因连接到PMD18-T质粒上,经酶切鉴定和序列测定证明该序列是PIL-6。序列分析结果表明:该基因cDNA全长741bp,开放阅读框由639个核苷酸组成,推测产生的编码产物由212个氨基酸组成。核酸序列分析比对发现:荣昌猪IL-6与GenBank已发表的IL-6序列的同源性较高,为99.8%。100%,氨基酸的同源性为99.5%-100%。对荣昌猪IL-6基因氨基酸的亲水性和蛋白表面可能性进行分析,表明其与IL-6基因氨基酸序列性质一致。The eDNA sequence of Rongehang porcine interleukin -6( PIL -6) was amplified by RT - PCR from totle RNA extracted from blood lymphoeytes, which were cultivated and stimulated with ConA. Then the amplified eDNA was cloned into PMD18 -T vector. IL -6 was proved by enzyme digestion and sequence identification. The full length of the cloned eDNA ,was 741 bp with an ORF composed of 639 nueleosides,whieh eneedes 212 amino acids. It also showed there was high homology between the Rongehang PIL-6 and other IL-6 sequence published from the GenBank,whieh was an identity of99. 8% - 100% ,and the identity of amino acids was 99. 5% - 100%. Meanwhile ,the analysis of hydrophobieity ,antigenic index and surface probability analysis of PIL -6 protein were the same as the feature of the PIL-6 protein
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