Cloning And Expressing Of A Novel Human Gene NSP019

Objective Cloning and expressing of a novel human gene NSP019 to explore the possibility of a new drug.Methods Using bio-informatics computer software Electronic-Signal Sequence Trap (ESST) technique, we analyzed and predicted all the ESTs of a novel gene-NSP019. This gene was cloned by PCR from cDNA library of pituitary and was expressed in eucaryotic COS-7 cells and prokaryotic E.coli. By the methods of Western blot -. Northern blot and sub-cellular localization , further investigation was made to study the characterization of NSP019.Results Bio-informatics analysis showed that the whole length of NSP019 had 5716bp and ORF had 726bp , encoded 241-amino-acid which possessed an N-terminal signal peptide and a glutamine-rich region . Genomic structure showed that the gene included 5 exons and located on chromosome 2q32.We cloned this novel human gene by PCR and the ORF sequences of the gene was analyzed. Our results were coincidence with the information predicted by Bio-informatics . Northern blot showed that NSP019 was expressed widely in different tissues. Sub-cellular localization and Western blot indicated that NSP019 coded a secreting protein . NSP019 can also express in E.coli and the purified protein was achieved.Conclusion NSP019 is a novel human secreting protein (GenBank Accession NO. AF361495) . It may be play an important role in the regulation of major physiological processes, such as development, growth , metabolism , reproduction ,etc. The exact functions of the secreting protein remain to be studied .