Cloning and Expression Analysis of THE: a novel human gene high expression in testis

Select EST(CA423810) from UniGene bank.According to nr and EST database BLAST result,It was found CA423810 was a sequence that represents a new gene.This EST contig has a complete ORF.Length of The new gene complete cds was 2 232 bp,ORF was from 273 bp to 1493 bp.The gene is located in 5q22 coding a 44 859 Dalton protein composed of 388 amino acid.RT-PCR was used to detect the expression level of the new gene which was cloned by using EST contig.Eleven samples of tumor and corresponding normal tissues,Human testis cDNA library and Human embryo brain cDNA library were adopted in this study.Firstly,Total d from the sample of tumor and corresponding normal tissues.cDNA was synthesized by using Reverse Transcription System.Then,PCR amplification was performanced by using the gene specific primers designed according to the new gene cDNA sequence with tissues cDNA and cDNA library as the templates respectively.In the mean while,the house-keeping gene GAPDH be used as comparision to test the expression level of mRNA.Our research suggested that the new gene only expressed in testis tissue highly.In other tumor tissues and corresponding normal tissue no gene’s expression was detected obviously.Therefore,the new gene was named for testis high expression gene(THE).According to the result,we can conclude that the new gene THE is the isoform of the same gene expressed in Humam brain tissue.