摘要
根据G enB ank文献,应用O ligo 6.0分析软件设计合成了用于扩增鸭Ⅰ型干扰素(-αIFN)基因的2对引物,以麻鸭肝脏提取的基因组DNA为模板,采用N est-PCR方法扩增获得了约600 bp片段,经T载体克隆和测序分析证实,是麻鸭Ⅰ型干扰素基因(SD IFN-α)。生物信息学分析表明,SD IFN-α的开放阅读框架(ORF)由576个核苷酸所组成,预测蛋白质相对分子质量为21 640,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号氨基酸A和29号氨基酸S之间,序列中无内含子。成熟的多肽中含8个半胱氨酸和2个糖基化位点,2个蛋白激酶C磷酸化位点(prote in k inase C phosphory lation s ite)和2个酪蛋白激酶Ⅱ磷酸化位点(case in k inaseⅡphosphory lation s ite)。SD IFN-α与鸭α-IFN基因核苷酸同源性为99.3%,有4个碱基的差异;与北京鸭α-IFN基因的核苷酸同源性为99.1%,氨基酸同源性为98.96%;与鸡-αIFN的核苷酸同源性为71.8%。与鹅、狗、牛、马和人的-αIFN基因的核苷酸同源性分别为96.0%、45.8%、45.5%、44.6%和41.7%。遗传进化分析结果表明,IFN-α的这些同源性与其动物分类地位相一致。
Based on GenBank database, a pair of primer by Oligo6.0 software was abtained. Using Ma duck liver DNA as template, SDIFN-α gene about 600 bp was amplified by nest-PCR and then cloned in T carrier. Through sequencing and analyzing by method of bioinformatics： SDIFN-α ORF includes 576 nucleotides, encoding 191 amino acids. Molecular weight is about 21 640, the front 28 amino acide is signal peptide, and the cleavage site lies in 28-29 amino acide（ANA-FS）. There is no intron within the 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 with that of goose,dog,cattle,horse and human is 96.0% ,45.8% ,45.5%,44.6% and 41. 7% in nucleotide level. Phylogenetic development indicates： These homology of SDIFN-α are in accordance with its classification.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2006年第4期401-404,共4页
Chinese Journal of Veterinary Science
基金
国家科技攻关重大项目(2004BA901A03)
四川省重点建设学科资助项目(SZD0418)
关键词
麻鸭
α干扰素基因
克隆
序列分析
Ma duck
α-interferon gene
cloning
sequence analysis
