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《Journal of The Fourth Military Medical University》 2002-13
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Research of inducible expression of ndr2 gene in lung adenocarcinoma GLC-82 cells

LI Shu Jun 1,2 , WANG Hua 3, QI Hao Wen 2, ZHANG Jian 1, HE Peng 1 , ZHANG Wen Hong 1, JI Shao Ping 1, LIU Xin Ping 1, YAO Li Bo 1 1Department of Biochemistry and Molecular Biology, Faculty of Preclinical Medicine, 2Depar  
AIM To Construction a recombinant pIND ndr2 expression vector, to observe its steady expression in transfected human lung adenocarcinoma cell line GLC 82 and its role in further study of biological function of ndr2 for lung adenocarcinoma. METHODS ndr2 gene of human normal lung tissue and human lung adenocarcinoma cell line GLC 82 was amplified by PCR RT. PCR amplification was performed by using primers based on the known gene sequence of ndr2, and fragments of DNA was cloned into vector pUC19. The sequence of the fusion gene was analyzed. Ecdysone inducible mammalian expression system was used. A recombinant pIND ndr2 vector was constructed at its Hin dⅢ+ Eco RI sites. The recombinant pIND ndr2 vector and pIND vector were tranfected into GLC 82 cells respectively, and the cell clones were screened with G418 and zeocin. Expression of ndr2 gene was detected by RT PCR, and expression of ndr2 protein was detected by western blot and immunohistochemical method after the cells were induced with ponasterone A. RESULTS The sequence of ndr2 gene that was cloned into vector pUC19 was correct. ndr2 gene was correctly transfected into GLC 82 cells and was appropriately expressed with induction of panosterone A. The expression of ndr2 of lung adenocarcinoma cell GLC 82 transfected by pIND ndr2 with induction of panosterone A was high. CONCLUSION Recombinant pIND ndr2 expression vector was successfully constructed and expressed steadily in human lung adenocarcinoma cell line GLC 82.
【Fund】: 国家杰出青年科学基金资助 (3 982 5 113 ) ;; 国家重点基础研究发展规划 (973 )项目资助 (J19990 75 6
【CateGory Index】: R734.2
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