【作者】 陈斌； 程安春； 汪铭书； 余勇； 唐琼； 石谦； 张东； 彭健康； 陈海滨；

【Author】 Chenbin,Chenganchun,Wangmingshu,Yuyong,Tangqiong,Shiqian,Zhangdong,Pengjiankang,Chenhaibin (Key laboratory of animal disease and human health of Sichuan province, Avain disease research center of Sichuan Agricultural University, yaan,625014;Sichuan provincial animal epidemic prevention &.supervision general Office,chengdu,610041,Leshan city shawan animal husbandry bureau)

【机构】 动物疫病与人类健康四川省重点实验室,四川农业大学禽病防治研究中心； 四川省动物防疫监督总站； 四川乐山沙湾区畜牧局；

【摘要】 根据GenBank文献,应用Oligo 6．0分析软件设计合成了用于扩增鸭Ⅰ型干扰素(α-IFN)基因的两对引物,以麻鸭肝脏提取的基因组DNA为模板,采用Nest-PCR方法扩增获得了约600bp片段,经T载体克隆和测序分析表明获得了麻鸭Ⅰ型干扰素基因(SDIFN-α)。生物信息学分析表明, SDIFN-α开放阅读框架(ORF)由576个核苷酸组成,预测蛋白质分子量为21．64kDa,编码191个氨基酸,其中前28个氨基酸可能是信号肽部分,切割位点在28和29号氨基酸即ANA和FS之间,序列中无内含子。成熟的多肽中含8个半胱氨酸和两个糖基化位点,2个蛋白激酶C磷酸化位点(Protein kinase C phosphorylation)和2个酪蛋白激酶Ⅱ磷酸化位点(Casein kinase Ⅱ phosphorylation site)。 SDIFN-α与鸭α-IFN基因核苷酸同源性为99．3％,有四个碱基的差异;与北京鸭α-IFN基因核苷酸同源性为99．1％,氨基酸同源性为98．96％;与鸡α-IFN核苷酸同源性为71．8％。与鹅、狗、牛、马和人的α-IFN基因核苷酸同源性为分别为96％、45．8％、45．5％、44．6％和41．7％。遗传进化分析结果表明,IFN-α的同源性与动物分类地位是一致的。更多还原

【Abstract】 Based on Genebank database, we get a pair of primer by Oligo6.0 software. Using Ma duck liver DNA as template, SDIFN-α gene about 600bp was cloned by nest-PCR and then cloned in T carrier. Through sequencing and analyzing by method of bioinformatics: SDIFN-αORF includes 576 nucleotides, encoding 191 amino acids. Molecular weight is about 21.64kDa, front 28 amino acide is signal peptide, and the cleavage site lies in 28-29 amino acide(ANA-FS). There is intronless in gene. There includes 8 cysteine and 2 N-Glcosylation sites in mature peptides. Meanwhile there are two Protein kinase C phosphorylation and Casein kinase Ⅱ phosphorylation sites. The homology of SDIFN-α and duck interferon is 99.3% at nucleotide level. There are four nucleotides in difference. The homology of SDIFN-α and BDIFN-α is 99.1% in nucleotide level. The homology in amino acids is 98.96%. The homology of SDIFN-α and chicken Ⅰ interferon is 71.8% in nucleotide level. The homology of SDIFN-α and goose、dog、cattle、horse and human is 96%、45.8%、45.5%、44.6% and 41.7% in nucleotide level. Phylogenetic development indicates: The homology of SDIFN-αand the class is the same.更多还原