Chromosome Location and Elongation of Radiation-induced Expressed Sequence Tag by the Aid of Bioinformatics / 生物化学与生物物理进展

ABSTRACT

Regulation of gene expression is one of the most importa nt responses of cells to DNA damage induced by radiation. A novel expressed seq u ence tag (EST) fragment had been cloned from human embryo lung cells induced by 50cGy radiation and named RIG1. To clone the full-length cDNA of RIG1, a non-c loned cDNA library of human embryo lung cells induced by low dose irradiation ha d been established. This library was used as template in enchanced nest RACE PCR and biotin-labeled probe was used for further purification. The 3′ flanking s equence of this EST was cloned and sequenced with this set of technology. It was illuminated by homology analysis that this 3′ flanking sequence and the origin al EST are well aligned with a BAC clone of 20th chromosome and the predic ted exons sequence of this chromosome is well consisten ce with the real EST. Thus the RIG1 can be roughly located in 20th chromos ome. By use of the exons sequence predicted from chromosome sequence by GENSCA N, full-length of RIG1 gene has been cloned. Chromosome location of RIG1 gene i s further determined by this successful verification of Bioinformatics predictio n by experiment. By the same step, genome sequence of RIG1 has been determined. Therefore,by the combined use of Bioinformatics analysis，the full-length cDNA sequence and genome sequence of RIG1 gene are obtained and the predicted protei n sequence is determined.