The Study Of The Function Of HL52

For searching new hematopoietic control genes, we used mice bone marrow stromal cells and suppression substrictive hybridization to chose for the screening differentially expressed genese expressed by LTC cultured stromal cells but not by the uncultured bone marrow cells. We chosed one of novel genes which was mL52. The homologous gene of human was hL52 gene.In the former work we mainly studied the sequence and the structure of this gene, now in this paper we will explore the function of this gene and the biologic effect of the product of genetic expression. We used software, DNAStar, to analyse the structure of the product of genetic expression and according to the result we speculate that it may be one kind of antimicrobial peptides. To verify the speculation, we carry out the following experiments.1. Try to gain the HL52 through SDS-PAGE. E.coli BL21 which were transformed into vector pET22b (+) - hL52 (no tag) and then pick the amp+ transformants. Purified the vectors from the amp+ transformants and cleaved by enzymes to verify. Select three groups including positives, negatives and empty vector transformants and each group was separated to two parts in which one part was induced by IPTG and another was not induced by IPTG in the process of growth. Sometimes later centrifugated the culture and collected the deposition. Then use cell lysis solution to lyse E.coli BL21. And use the lysis products to make SDS-PAGE. In the result we did not find special bands which show the product of hL52 genetic expression would make effect as soon as expressed, so it was decomposed immediately and we can not see it in the SDS-PAGE.If it is antimicrobial peptide it will make effect on E.coli BL21, so we also can not find it. Whether the hypothesis is true, we will do the following experiments to verify it.2. Verify whether the HL52 is antimicrobial peptide or not. We cultivated the E.coli BL21 which were transformed into vector pET22b (+) - hL52 (no tag) in liquid media and induced by IPTG. If the growth of E.coli BL21 was inhibited, it was shown the product of hL52 genetic expression is antimicrobial peptide. There were several controls in experiments. The result of experiment showed the growth of E.coli...