Earticle

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

Abstract
 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

키워드

저자

• Hong Cao [ Department of Microbiology ]
• Liang Peng [ Department of Microbiology ]
• Zhenhua Ding [ Department of Radiation Medicine, School of Public Health and Tropical Medicine, ]
• Zhihua Liu [ Department of Infectious Diseases, Nanfang Hospital, Southern Medical University ]
• Xiaolu Liu [ Department of Microbiology ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  International Journal of Advanced Science and Technology

• 간기

  월간

• pISSN

  2005-4238

• 수록기간

  2008~2016

• 십진분류

  KDC 505 DDC 605