Carcinogenesis, Teratogenesis & Mutagenesis ›› 2002, Vol. 14 ›› Issue (3): 171-173.doi: 10.3969/j.issn.1004-616x.2002.03.009

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CONSTRUCTION AND IDENTIFICATION OF STABLE EXPRESSION SYSTEM OF KARYOCYTES WITH TEF-1δ GENE

LEI Yi-xiong1, Pius Joseph2, CHEN Jia2kun1, Ong Tong-man2   

  1. 1. Institute for Chemical Carcinogenesis, Guangzhou Medical College, Guangzhou 510182, P.R. China. 2. Molecular Epidemiology Laboratory, Toxicology and Molecular Biology Branch National Institute for Occupational Safety and Health, Morgantown, WV, USA
  • Received:2002-03-21 Revised:2002-05-14 Online:2002-07-30 Published:2002-07-30
  • Contact: LEI Yi-xiong

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Abstract: Purpose : To const ruct and identify the stable expression system of karyocytes with TEF2δ gene. Methods : Two stable t ransfections of CHO and COS7 cells with plasmid (pcDNA3. 1/ V52His2TOPO Vector) expressing TEF21δcDNA were established by using calcium phosphate and G418 selection protocols. Results : The result s showed that , after G418 selection and western blotting analysis , 3 out of 10 CHO cell lines t ransfected with TEF21δcDNA expressed very high levels of TEF21δencoded protein with an approximately molecular weight of 31 kDa. as compared with vector cont rol t ransfectant s that showed no expression , and compared with the other cell lines that expressed relatively low proteins. Similarly , 4 out of 4 COS7 cell lines had significant overexpression of TEF21δencoded protein. The names of these stable t ransfection cell lines were CHO2pcDNA3. 12TEF21δ, # 3 , # 6 and # 14 as well as COS72pcDNA3. 12TEF21δ # 4 , # 8 , # 14 and # 17 , respectively. Conclusion : These cell lines can be applied to functional studies of the TEF21δ gene , and these are the optimal cell lines for studies on the underlying molecular carcinogenic mechanisms of Cd carcinogenesis.

Key words: TEF-1δgene, Stable transfection, CHO cells, COS7 cells, Western Blot

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