文章摘要
Ying Shi,Shigang Cheng,Xu Chen,Chuanguo Xiao.[J].南京医科大学学报,2008,28(5):304~307
Construction and identification of recombinant lentivirus-mediated gene transfer system for rat transducer of regulated CREB activity 1
投稿时间:2008-08-20  
DOI:10.7655
中文关键词: 
英文关键词: TORC1  lentivirus vector  spinal cord injury
基金项目:
作者单位
Ying Shi Department of Urology, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science & Technology,Wuhan 430022, Hubei Province, China 
Shigang Cheng  
Xu Chen  
Chuanguo Xiao  
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中文摘要:
      
英文摘要:
      Objective: To construct a recombinant lentivirus vector which carries SD rat transducer of regulated CREB activity-1(TORC1) gene and examine its ability to express the TORC1 gene in vitro. Methods: The coding sequence of SD rat TORC1 gene was amplified using PCR and cloned into pGC-FU vector. 293T cells were transfected using Lipofectamine 2000 and packaged for the recombinant lentivirus particles. When the cloned sequence was identified to be right, the recombinant lentivirus particles were amplified in a large quantity. The titer of virus was determined by real-time PCR and the level of TORC1 expression was examined by Western blot. Results: The recombinant lentivirus vector carrying TORC1 was constructed successfully and could express TORC1 at a high level in 293T cells in vitro, and the titer determined by real-time PCR was 2×108 TU/ml. Conclusion: The recombinant lentivirus vector could express TORC1 gene at a high level, and was very helpful in the study of exploring the effect of TORC1 on spinal cord injury.
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