中文摘要：

目的：构建含有小鼠趋化因子受体-7（CCR7）基因的重组腺病毒，为下一步转染不成熟树突状细胞（imDC）诱导免疫耐受研究奠定基础。方法：提取小鼠胸腺总RNA，应用逆转录PCR（RT-PCR）方法，以自行设计的带有酶切位点的引物，扩增获得CCR7基因全部序列，经过T-A克隆，酶切亚克隆到穿梭质粒pAdTrack-CMV上，在BJ5183菌内和pAdeasy-1同源重组，筛选阳性克隆，酶切鉴定，线性化后脂质体法转染HEK293细胞进行包装、PCR鉴定及扩增，得到含有CCR7基因的重组腺病毒，根据报告基因GFP测定病毒滴度。结果：成功构建小鼠CCR7基因重组腺病毒，病毒滴度为1×10^9U／mL。结论：该重组腺病毒载体的成功构建，为进一步研究携带CCR7基因的imDC的趋化迁移性等研究提供了一定的工作基础。

英文摘要：

Objective： To construct the recombinant adenovirus carrying mouse CCR7 gene for further study： Methods： The CCR7 gene was amplified by RT-PCR from thymus of mouse, after sequencing, the fragment was cloned into T vector and subcloned into pAdTrack-CMV. The linearized shuttle plasmid was homogenously recombined with pAdeasy-1 in E. coli B J5183. The candidate clone was further analyzed by restriction endonuclease digestion. Then the recombined plasmid was transfected into HEK293 cells for packaging, PCR identification and amplifying, Infection titer was monitored by green fluorescent protein （GFP） expression. Results： The mCCR7 gene was cloned into the adenovirus successfully. Viral titer checked by GFP was about 1 ×10^9U/mL. Conclusions： The constructed recombinant adenovirus containing mCCR7 lays a potent foundation to investigate the chemotaxis ofimDC transfected by mCCR7.