In Silico Cloning Partial ESTs From CDNA Library Of Normal Swine Peripheral Blood Lymphocytes And Experimental Identification

Recently, most of swine genes obtained by silico cloning were related to its important economic characters, such as color of hairs, lipid metabolism, meat quality, reproduction ability, and so on. However, the genes associated with innate immunity and adaptive immunity were seldomly reported. In this study, we attempted to find a good way to get partial or full-length genes by silico cloning through bioinformatic databases in order to obtain novel genes related to influencing viral reproduction and anti-virus immune response fastly.We carried out silico cloning partial ESTs from cDNA library from normal swine peripheral blood lymphocytes. 20 contigs of them were assembled into 20 clusters. They conta i ned full genes ORFs and proved to have been prolongated effectively. The result showed that the seqences of 14 genes were genes known previously and 6 genes were new .We separately chose one seqence from known and the other from unknown genes to make full-length cDNA annotation. Their code number was EST contig110 and EST contig133. We designed specific primers to amplify the two genes by RT-PCR, and then sequenced. The result was obtained as expected and proved that silico cloning genes was reliable. We fuether predicted physical and chemical property, hydrophobicity and secondary structure of a novel swine protein, 60S ribosomal protein L18a. In human being, this protein could interact with the internal ribosomal entry sequence (IRES) of HCV, acted as key roles during the process of viral protein translation.It was worthy further study...