A combination of biotin labeled screeing of cDNA library and RACE technique to identify a novel human gene HRNT-1

Objective： An EST fragment, Rat ZA73 (GenBank Accession Number： AF011363 ), was cloned by using mRNA differential display technique from rat tracheal epithelial (RTE) neoplastic transformed cell model induced by α particles radiation. Homology search in GenBank showed that rat ZA73 was a novel EST. Based on the previous work, this study is to identify human full length sequence of EST. Methods： According to the sequence of rat ZA73, a biotin labeled probe was used to screen a human cDNA library, and then the gene sequence was extended by RACE (rapid amplification of cDNA ends). Results： Human gene named HRNT 1 (GenBank Accession Number： AF223393) was sequenced. Its total length is 4256 bp in length, with an ORF located in the region between 254 and 3013 bp. 5′ UTS (untranslated sequences) is 253 bp, 3′ UTS is 1240 bp. Conclusions： The combination of biotin labeled screening of cDNA library and RACE is an effective method to clone full length cDNA, especially for those with sequence longer than 2 kb.