【摘要】 干扰素（ＩＦＮ）是一种具有广谱抗病毒、抗肿瘤以及免疫调节等活性的细胞因子。本研究通过ＰＣＲ从ＡＡ肉鸡基因组ＤＮＡ中克隆了ＩＦＮ－ａ（ＣｈＩＦＮ－ａ）基因，测序分析，并将该基因与表达载体ｐＱＥ３０相连，构建重组表达质粒ｐＱＥ３０／ＣｈＩＦＮ－ａ，以ＩＰＴＧ诱导在Ｍ１５中进行表达。测序结果表明ＣｈＩＦＮ－ａ的开放阅读框（ＯＲＦ）是由４８６个核苷酸编码的，成熟蛋白为１６２个氨基酸，有４个糖基化位点，推测分子量为１９ｋＤ。与ＤＤＢＪ／Ｇｅｎｂａｎｋ中鸡ＩＦＮ－ａ基因序列一致。表达产物经ＳＤＳ－ＰＡＧＥ分析、ＩＤ－ＭＵＬＴＩ分析及Ｗｅｓｔｅｍ　Ｂｌｏｔ分析表明，表达出１９ｋＤ左右的融合蛋白，表达的蛋白量占总菌体蛋白的６．５％，包涵体占沉淀蛋白的３０％。本研究为重组干扰素用作新型广谱抗病毒生物制剂和免疫增强剂的开发奠定了基础。更多还原

【Abstract】 Interferons (IFN) represent a family of cytokines that share the capacity to inhibit viral replication and to exert effects on immune function. In this research interferon-a (IFN-a) gene was cloned from genomic DNA of Arbor Acres chicken (AA chicken) by polymerase chain reaction (PCR), and was ligated to pGEM T Easy vector for sequencing. This gene was then digested from pGEM T-Easy vector/ChIFNa by Sph I and Hind Ⅲ and legated to high level expression vectors pQE30 to construct expression plasmids poE30/ChIFN-a. Sequencing results show that the ORF of the cloned ChIFN-a gene is composed of 486 nucleotide which encoding 162 amino acid and showing no difference with ChIFN-a gene from DDBJ/Genbank. M15 strains with pQE30/ChIFN-a can produce the protein with the molecular weights of 19 kD on SDS-PAGE, which are consistent with the putative molecular weightS of lChIFN-a. Western blot analysis proves that the protein expressed by M15 is the special recombinant ChIFN-a.更多还原