关键词: cDNA末端快速扩增, 反式二羟环氧苯并(a)芘, 基因

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