| 杨阳 郑智坤 马骥 杜成 郭燕 刘文超.非小细胞肺癌ROS1及其融合基因CD74-ROS1逆转录病毒载体的构建[J].,2015,15(13):2440-2442 |
| 非小细胞肺癌ROS1及其融合基因CD74-ROS1逆转录病毒载体的构建 |
| Construction of the Retroviral Vectors Carrying the ROS1 and CD74-ROS1Fusion Gene in Non-small Cell Lung Cancer |
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| DOI： |
| 中文关键词: 逆转录病毒载体 lNSCLClROS1l 融合基因 |
| 英文关键词: Retroviral vector NSCLC ROS1 Gene-fusion |
| 基金项目:国家自然科学基金项目（31371375） |
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| 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Ham H I和Taq I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
载体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 Taq I and Bam H I, 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.Conclusion: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. |
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