Carcinogenesis, Teratogenesis & Mutagenesis ›› 2006, Vol. 18 ›› Issue (4): 302-305.doi: 10.3969/j.issn.1004-616x.2006.04.013

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Full-length Cloning of brg,the Anti-BPDE Related Gene

AN She-juan1,3, CHEN Jia-kun2,, CHEN Hua-jie1, CHANG Wei1,LIU Li-li2, ZHAO Yan-feng2, CHEN Xue-min1   

  1. 1.Department of Occupational and Environmental Health, School of Public Health,Tongji Medical College,Huazhong University of Science and Technology,Wuhan Hubei 430030,China; 2.Institute of Chemical Carcinogenesis,Guangzhou Medical College, Guangzhou 510182, Guangdong; 3.Lung Cancer Institute, Guangdong Provincial People's Hospital, Guangzhou 510080, Guangdong,China
  • Received:2005-05-27 Revised:2005-12-19 Online:2006-07-30 Published:2006-07-30
  • Contact: CHEN Jia-kun

Abstract: BACKGROUNG & AIM: Previous studies showed a gene fragment was identified as being over-expressed in 16HBE-C cells when compared to 16HBE cells, which is named brg (anti-BPDE related gene). The aim of this study was to obtain the full-length cDNA of brg based on its EST fragment and to provide basic data for the functional research of the novel gene. MATERIAL AND METHODS: Based on the rapid amplification of cDNA ends (RACE) method, gradient and nested PCR protocols were used to obtain full-length cDNA in order to get a discrete and bright band on an agarose gel electrophoresis. The nested PCR products were then sequenced and the bioinformatic analysis was carried out. RESULTS: Full-length cDNA of the gene was successfully cloned by both 3'-end RACE and 5'-end RACE. The cDNA had a total length of 1 214 bp. Sequence analysis of the full-length cDNA of the gene revealed that the cDNA encoded an open reading frame of 1 032 bp; with the deduced protein containing 344 amino acids. CONCLUSION: The general and simple protocol was useful for cloning of full-length gene based on EST fragment. The results suggest that brg gene may be involved in carcinogenesis during anti-BPDE related tumor development.

Key words: rapid amplification of cDNA ends, anti- benzo [a]pyrene -7, 8-diol -9, 10-epoxide, gene

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