中文摘要：

目的 构建表达转化生长因子βⅡ型受体（TβRⅡ）胞外区一活化T细胞表达和分泌的调节因子（R-ANTES）融合基因重组腺病毒并鉴定其在肿瘤细胞的表达。方法 采用逆转录-聚合酶链式反应（RT-PCR）方法扩增小鼠TβRⅡ胞外区和RANTES基因，重叠PCR法扩增TβRⅡ胞外区-RANTES融合基因，将融合基因克隆至pDC316载体，采用AdMax Adenovirus Vector Creation试剂盒构建融合基因重组腺病毒载体，融合基因重组腺病毒按moi=100：1的比例体外感染小鼠肺腺癌L795细胞系，Western Blot方法检测感染后肿瘤细胞的蛋白表达。结果 RT-PCR正确扩增了440bpTβRⅡ胞外区、244bp RANTES基因，重叠PCR正确扩增了745bpTβRⅡ胞外区-RANTES融合基因，重组质粒融合基因-pDC316经酶切鉴定正确，与pJM17双质粒共转染获得表达融合基因重组腺病毒，病毒滴度为8×10^10pfu／ml，感染后的LA795细胞有相对分子质量为33000的蛋白表达。结论 成功构建表达TβRⅡ胞外区-RANTES融合基因重组腺病毒载体，为进行体内抗肿瘤基因治疗的研究创造条件。

英文摘要：

Objective To construct a recombinant adenovirus vector containing extracellular domain of TβRⅡ -RANTES fusion gene and identify its expression. Methods Mouse origin extracellular domain of TβRⅡ and RANTES gene were amplified by RT-PCR method; Extracellular domain of TβRⅡ- RANTES fusion gene was amplified by overlapping PCR method; Extracellular domain of TβRⅡ-RANTES fusion gene was cloned into pDC316 vector,and recombinant adenovirus vector containing the fusion gene was constructed by the AdMax Adenovirus Vector Creation System;Recombinant adenovirus was transfected into LA795 cells, and the expression of recombinant adenovirus was detected by Western blot. Results Extracellular domain of TβRⅡ and RANTES gene were amplified by RT-PCR; Extracellular domain of TβR Ⅱ -RANTES fusion gene was amplified by overlapping PCR; Recombinant adenovirus vector containing fusion gene was constructed successfully and its titer was 8 × 10^10 pfu/ml;33 000 fusion proteins was detected in LA795 cells infected with recombinant adenovirus containing fusion gene. Conclusion Recombinant adenovirus vector containing extracellular domain of TβRⅡ -RANTES fusion gene was constructed successfully, and it may be used as a novel agent for gene therapy in anti-tumor.