鸡白细胞介素2的克隆及其序列分析  

Cloning and Sequence Analysis of Chicken Interleukin-2

作　　者：白程乐 申艳[1] 高冬妮[1] 于航[1] 平文祥[1] 葛菁萍[1] 

机构地区：[1]黑龙江大学生命科学学院/微生物省高校重点实验室,哈尔滨150080

出　　处：《中国农学通报》2017年第26期117-122,共6页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金"表达IBDV主要保护性抗原的重组杆状病毒构建及其作为疫苗的免疫效果研究"(31270143);黑龙江省高等学校科技创新团队"农业微生物发酵技术"(2012td009)

摘　　要：旨在从分子生物学角度进一步研究鸡免疫刺激因子白细胞介素2(IL-2),探究IL-2蛋白结构。根据GenBank发表的IL-2序列设计引物扩增IL-2基因,将其克隆到p GM-T载体后进行鉴定,鉴定正确的pT-IL-2重组质粒,然后对其进行序列分析。应用生物信息学软件对IL-2序列进行跨膜区、磷酸化位点、氨基酸序列、开放阅读框进行分析和IL-2蛋白的三级结构进行预测。核苷酸序列测定结果表明:IL-2基因片段为430 bp,编码143个氨基酸。生物信息学分析结果表明:本实验获得的IL-2基因编码的蛋白质中含有1个丝氨酸和3个苏氨酸,这可能成为蛋白激酶磷酸化的位点。IL-2蛋白可能在5~28位含有跨膜区。其蛋白二级、三级结构预测结果显示其属于亲水性蛋白,多为α-螺旋、β-转角和N端无规卷曲结构。该结果为IL-2的分子生物学探究奠定了基础。We aimed to study the immune stimulating factor IL-2 from the perspective of molecular biology and explore its protein structure.Primer 5.0 was used to design a pair of primers according to the sequence of IL-2 in GenBank.IL-2 gene was amplified and cloned to p GM-T vector,and identified by restriction enzyme,and PCR.Then the nucleotide sequence was analyzed.Bioinformatics software was used to analyze the amino acid sequence,transmembrane domain,phosphorylation sites,and predict the secondary and tertiary structure of IL-2.The results revealed that IL-2 gene was 430 bp and encoded 143 amino acids.Bioinformatics analysis indicated that IL-2 protein contained 1 serine and 3 threonines,which might become phosphorylation sites of protein kinase.Besides,IL-2 might contain transmembrane domain within sites 5-28.The secondary and tertiary structure prediction results indicated that IL-2 was a hydrophilic protein,and dominated byα-helix,β-turn and random coil at N terminal.The study lays a foundation for the molecular research of IL-2.

关 键 词：白细胞介素2 基因克隆 序列分析 蛋白结构预测 

分 类 号：Q786[生物学—分子生物学]