원문정보
초록
영어
Scanning, comparing and designing of candidate siRNAs to HBV genes have been performed by searching he DNA databases and using network servers. The homology search among the RNAi targeted region on RNA sequence of HBV and human genome was done with BLAST. Three candidate siRNAs to HBV were chosen to construct three shRNA expression vectors respectively. Two complementary DNA oligonucleotides linked by 9-nucleotide loop and RNA polymerase Ⅲ terminators were designed first, and the annealed hairpin siRNA was inserted into the plasmid that carries green fluorescent protein. Then these recombinant plasmids were transformed in Escherichia Coli and detected by PCR and sequencing. Finally, the plasmids were used to transfect HeGp2 cells with liposome. The results showed the molecular weight of PCR products matched predicted size, and three siRNA plasmids were sequenced as expected sequences. The transfected cells stained with green fluorescence were also observed. Thus the shRNA expression plasmid specific for interfering HBV replication was successfully constructed. Keywords: RNAi; siRNA; HBV; bioinformatics; computer scanning.
목차
1. Introduction
2. Materials and Methods
3. Results
3.1 Bioinformatics and computer-based design of siRNA to HBV
3.2 DNA oligonucleotides sequences design
3.3 Identification of recombinant siRNA plasmid
3.4 Transfection of HepG2 cell with recombinant siRNA plasmid
4. Discussion
References