ABSTRACT
BACKGROUND:MicroRNAs (miRNA) through regulating specific target gene mRNA expression play important roles in different processes of diseases. OBJECTIVE:To study the interaction of miRNA-21 with its target gene TLR4 in Hela cels. METHODS:The candidate target gene of miRNA-21 was determined according to miRNA analysis databases. The constructed recombinant adenovirus vector carrying pri-miRNA-21 gene was used, which could package and amplify viruses to transfect Hela cels. Then, the expression of fluorescent proteins was detected. Forty-eight hours after transfection of miRNA-21 or control, extracted proteins were used for detection of TLR4 protein using western blot assay. RESULTS AND CONCLUSION:Recombinant adenoviruses pAd/pri-miRNA-21 and pAd/neg at 100 MOI could successfuly infect Hela cels. Bioinformatic analysis suggested several possible binding sites between miRNA-21 and TLR4. The experimental results showed that miRNA-21 down-regulated TLR4 at protein levels, indicating that miRNA-21 can interfere with the expression of TLR4 target gene.
ABSTRACT
A recombinant plasmid containing cathepsin B endopeptidase of Schistosoma japonicum(Sjcb2)was constructed,indentified by PCR,restrictive enzyme,digestion and DNA sequencing,and expressed into mammalian cells.Immunochemistry examination showed that the Sjcb2 gene can be expressed in the eukaryotic system,providing a basis for the development of schistosome DNA vaccine.