中文摘要：

目的构建结核分枝杆菌小分子热休克蛋白Hsp16.3原核表达载体,获得Hsp16.3蛋白,并应用生物信息学软件,分析结核分枝杆菌小分子热休克蛋白16.3（Hsp16.3）的多种蛋白质性质,以及获取该基因的相关信息。方法以结核分枝杆菌H37Rv株基因组DNA为模板,应用PCR方法扩增Hsp16.3基因,PET28a为载体,构建重组质粒PET28a-Hsp16.3并表达,利用Pmtparam、Protseale、HMMTOP、TMHMM、GOR4、ProtFun等软件预测和分析结核分枝杆菌小分子热休克蛋白16.3的氨基酸组成、理论等电点、结构域、活性位点、疏水性、跨膜区、糖基化位点、磷酸化位点、二级结构及系统进化分析。结果成功构建了Hsp16.3原核表达载体PET28a-Hsp16.3,并在宿主菌BL21（DE3）中表达。推测结核分枝杆菌小分子热休克蛋白16.3白分子式为C716H1131N197O225S4,分子量为16227.2,理论等电点为5.00。该蛋白氨基酸序列中含有α-螺旋（Alpha helix）46处,延伸链（Extended strand）25处,无规则卷曲（Random coil）73处及5个磷酸化位点;4个O-糖基化位点;1个前肽裂解预测位点。结论 Hsp16.3基因克隆入宿主菌中并成功表达;该蛋白推测是一个亲水性不稳定蛋白,无跨膜区,可能参与重要的能量代谢及免疫反应。

英文摘要：

[Objectives] To construct small heat shock protein 16.3 （Hsp 16.3） prokaryotic expression vector of Mycobacterium tuberculosis, obtain information of Mycobacterium tuberculosis small heat shock pro- tein 16.3 （Hsp 16.3）, and analyze the character of this gene and its product protein by a series of bioinfor- matics software. [Methods] The Hspl6.3 gene was amplified from Mycobacterium tuberculosis H37Rv genom- ic DNA by PCR.~The PCR products were further inserted into expression vector PET28 ct to construct the re- combinant plasmid PET28 ct- Hspl6.3, then transformed into E,coli BL21 （DE3） for expression. The character of Hspl6.3 protein such as the constitution of amino acid, equipotential, domain, active site, coiled coil, bydrophobicity, glycosylation sites, phosphorylation sites and second structure were analyzed by the Pmtparam, Protseale, HMMTOP, TMHMM, GOR4 and ProtFun, [Results] The Hspl6.3 was constructed and expressed in BL21 （DE3） successfully. Theoretical pI of this protein was 5.00, and the formula wag C716Hl131N1970225S 4. The results also showed that Hspl6.3 protein had 46-helix, 25 extended strand, 73 Random coil, 5 phos- phorylation sites, 4 glycosylation sites and a former peptide cracking prediction site. [Conclusion] The target gene （Hspl6.3） has been cloned into the host bacterium and expressed successfully. It is a hydrophilic and unstable protein without transmembrane area, and may participate in important energy metabolism and immune response.