胞内分枝杆菌HBHA基因的克

发布时间：2018-11-07 08:29

【摘要】：【目的】克隆胞内分枝杆菌肝素结合血凝素(HBHA)基因,并对其进行生物信息学分析和原核表达。【方法】应用巢式PCR克隆胞内分枝杆菌HBHA基因,利用在线分析软件对其基因序列及编码蛋白序列进行生物信息学分析。构建原核表达载体pET32a-HBHA,并进行诱导表达。【结果】胞内分枝杆菌HBHA基因完整的ORF全长618bp,编码205个氨基酸,该基因氨基酸序列与M.avium ATCC 25291有较高的相似性。HBHA蛋白为碱性、亲水性蛋白质,含12个潜在的磷酸化位点,存在抗原表位,亚细胞定位主要存在于细胞核,蛋白空间结构以α-螺旋和无规则卷曲为主。构建了pET32a-HBHA原核表达载体,其可在原核表达系统中诱导表达大小约为38ku的HBHA重组蛋白。【结论】胞内分枝杆菌HBHA是一种抗原指数较高的碱性、亲水性蛋白,可在原核表达系统中被表达。
[Abstract]:[objective] to clone and analyze the (HBHA) gene of Mycobacterium Intra-cell heparin binding hemagglutinin (HBHA) by bioinformatics analysis and prokaryotic expression. [methods] the HBHA gene of Mycobacterium endocytogenes was cloned by nested PCR. Bioinformatics was used to analyze the gene sequence and the coding protein sequence with online analysis software. The prokaryotic expression vector pET32a-HBHA, was constructed and induced to express. [results] the complete ORF of Mycobacterium HBHA gene was 618 BP, encoding 205 amino acids. The amino acid sequence of this gene is similar to that of M.avium ATCC 25291. The HBHA protein is alkaline and hydrophilic. It contains 12 potential phosphorylation sites and has antigenic epitopes. The spatial structure of protein was mainly 伪-helix and irregular curl. The prokaryotic expression vector of pET32a-HBHA was constructed, which could induce the expression of HBHA recombinant protein about the size of 38ku in the prokaryotic expression system. [conclusion] HBHA is a basic and hydrophilic protein with high antigen index. It can be expressed in prokaryotic expression system.
【作者单位】：吉林农业大学动物科学技术学院;长春理工大学生命科学技术学院;
【基金】：国家科技支撑计划项目(2007BAD55805) 国家自然科学基金项目(31272566) 吉林省科技发展计划项目(20120235)
【分类号】：R186

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