荣昌猪白细胞介素-6基因的克隆与序列分析

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(ConA)诱导猪淋巴细胞并提取总RNA,用RT-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741 bp,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。

Cloning and sequencing of IL -6 from Rongchang porcine

The cDNA sequence of Rongchang porcine interleukin6(PIL-6) was amplified by RT-PCR from totle RNA extracted from blood lymphocytes,which were cultivated and stimulated with ConA.Then the amplified cDNA was cloned into PMD18-T vector.IL-6 was proved by enzyme digestion and sequence identification.The full length of the cloned cDNA,was 741 bp with an ORF composed of 639 nucleosides,which encodes 212 amino acids.It also showed there was high homology between the Rongchang PIL-6 and other IL-6 sequence published from the GenBank,which was an identity of 99.8%~100%,and the identity of amino acids was 99.5%~100%.Meanwhile,the analysis of hydrophobicity,antigenic index and surface probability analysis of PIL-6 protein were the same as the feature of the PIL-6 protein.