中文摘要：

目的构建人热休克蛋白B6（HspB6）重组表达载体,初步研究其生物学活性。方法采用RT-PCR法扩增HspB6CDS段序列,并将扩增产物与小鼠Fc片段融合成重组基因HspB6/mFc,测序正确后插入蛋白表达载体pCEP4,随后转染人肾上皮293T细胞。扩增培养后收集细胞上清液用于浓缩、纯化以及Western blot验证。采用CCK-8法和Transwell实验分别检测HspB6/mFc融合蛋白对人脐静脉内皮细胞（HUVEC）增殖和迁移的影响。结果成功构建pCEP4/HspB6/mFc重组载体,并获得稳定表达HspB6/mFc融合蛋白的293T细胞。HspB6/mFc融合蛋白能明显促进HUVEC增殖和迁移。结论 pCEP4/HspB6/mFc重组载体及稳定表达的HspB6/mFc融合蛋白为进一步研究HspB6的生物学功能提供物质基础。

英文摘要：

Objective To construct a recombinant expression vector carrying human heat shock protein B6（HspB6） and preliminarily investigate its biological activity. Methods The CDS region of HspB6 was amplified by RT-PCR, which was fused with Fc fragment of mouse to construct recombinant HspB6/mFc gene. After identification by sequencing, HspB6/mFc was inserted into pCEP4 expression vector and then transfected into human embryo kidney 293T cells. The cell supernatant was collected for concentrating, purifying and confirming by Western blot. The effect of HspB6/mFc fusion protein on the proliferation and migration of human umbilical vein endothelial cells （HUVEC） was detected by CCK-8 and Transwell assay, respectively. Results The recombinant pCEP4/HspB6/mFc vector and its transgenic 293T cells stably expressing HspB6/mFc fusion protein were successfully constructed. HspB6/mFc fusion protein could significantly promote HUVEC proliferation and migration in vitro. Conclusion The recombinant pCEP4/HspB6/mFc vector and 293T cells stably expressing HspB6/mFc fusion protein pave the way for further study on biological function of HspB6.