中文摘要：

目的：利用逆转录病毒载体pBaba-puro构建携带ROS1基因及其CD74-ROS1融合基因的重组载体pBaba-puro-ROS1，pBaba-puro-CD74-ROS1。方法：设计与合成引物，提取组织标本RNA，反转录和PCR扩增，经BamHI和TaqI双酶切，琼脂糖凝胶电泳，切胶回收进行连接转化，并再次酶切鉴定，测序分析。结果：成功构建携带ROS1基因及其CD74-ROS1融合基因的重组栽体pBaba—puro-ROS1，pBaba—puro—CD74-ROS1，并通过双酶切与测序鉴定。结论：利用逆转录病毒载体基因重组技术能够成功构建出携带相应基因的逆转录病毒，可用于后续研究。

英文摘要：

Objective： Use pBaba-puro retroviral vetor to construct recombinant vector which carrying ROS1 gene and CD74-ROS1 fusion gene. Methods： Primers was designed and synthetized. Extracted tissue RNA, then apply it to reverse transcription for the template of latter PCR amplication. After PCR amplication and digested by TaqI and BamHI, agarose gel electrophoresis was used to detect the correct fragments to connect by T4 ligase. By E.coli transformation, restriction enzyme digest again to verify, then bring the right one to sequence. Results： The vector carrying ROS1 and CD74-ROS1 was successfully constructed. The verification of double digestion and sequencing was fine. Conclusions： The use of recombinant retrovirus vector technology can successfully construct a retrovirus carrying the corresponding gene which can be used for the following-up study.