Molecular Cloning, Bio-information Analysis And Preliminary Study On The Function Of ALT04ag Gene Associated With Human Lung Carcinoma Antigen

ALT-04, a specific monoclonal antibody against the human lung carcinoma was found by our laboratory. The cDNA fragment of ALT04ag, the ALT-04 related antigen gene, was cloned and proved that it was a new cDNA sequence with some extent transforming activities. In this paper the full length of ALT04ag cDNA was further cloned .The structure and function of this gene was also analyzed in order to provide soma new data for understanding the mechanism of the carcinogenesis and development of lung cancer.1. The full length of ALT04ag cDNA was further cloned by 5' RACE reaction. The homology analysis, molecular weight, transmembran domain, functional binding sites, isoelectric point, secondary structure and epitope of the protein coded by ALT04ag were analyzed and predicted by bioinformatics. There are 1115bp in ALT04ag cDNA. It coded a protein of 194 amino acids and probably a transmembran protein. Homology analysis showed that it was similar to AF219116 cDNA in some degree.2. The proliferation rate of the cells transfected by recombinant ALT04ag expression plasmid increased and these transfectant possessed tumorigenicity in nude mice, meanwhile the ALT04ag gene expression was detected and c-myc gene expression was upregulated.3. The expression plasmid of antisense ALT04ag cDNA was also constructed and transfected to the human lung squamous carcinoma cells L78 .The transfected cells showed growth inhibition and lost the tumorgenic property. Gene chip analytical results showed that the antisense ALT04ag cDNA could down regulate the proliferation associated genes, up regulated the growth inhibition gene and effect...