中文摘要：

目的获得编码VEGF189蛋白的基因,并通过原核表达系统表达VEGF189融合蛋白。方法应用SOE PCR技术合成VEGF189基因,克隆入pET-32a(+)原核表达载体,转化E.coliBL21(DE3)感受态细胞,IPTG诱导表达,进行SDS-PAGE和Western blot检测,表达产物经HisTrapTMHP亲和柱层析纯化,测定蛋白浓度。结果DNA测序分析表明,合成的VEGF189基因与文献报道相一致。在大肠杆菌BL21(DE3)中表达的Trx-VEGF189融合蛋白,以包涵体和可溶2种形式存在。表达产物可被兔抗鼠VEGF多抗特异识别。经HisTrapTMHP亲和柱纯化,获得纯度达85%的融合蛋白。结论成功合成VEGF189基因,并在原核表达系统中获得高表达,为研制高效抗肿瘤的VEGF抗体和疫苗奠定了前期基础。

英文摘要：

Objective To synthesis the gene for encoding VEGF189 and express VEGF189 protein in E.coli.Methods VEGF189 gene was synthesized using SOE PCR and cloned into pET-32a(+)vector.Then the recombinant expression plasmids were transfected into E.coli BL21(DE3)for expressing the fusion protein Trx-VEGF189. The expression products were identified by SDS-PAGE and Western blotting analysis respectively,and purified by HisTrapTM HP chromatography.Results DNA sequencing showed that VEGF189 gene was obtained successfull...