Design of siRNAs and Construction of Plant Expression Vector for Interfering with Tobacco Mosaic Virus
- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF APPLIED & ENVIRONMENTAL BIOLOGY
- Issue:
- 2
- DOI:
- 10.3321/j.issn:1006-687X.2006.02.002
- Key Word:
- RNA干扰(RNAi);发卡RNA(hpRNA);小分子干扰RNA(siRNA);烟草花叶病毒(TMV);植物表达载体;RNA interference (RNAi);hairpin RNA (hpRNA);short interfering RNAs (siRNAs);tobacco mosaic virus (TMV);plant expression vector
Abstract： RNA interference (RNAi) is now established as a powerful tool to silence gene expression in a variety of organisms. Depending on the gene sequence of tobacco mosaic virus ( TMV), 3 cDNA sequences that could generate short hairpin RNA were chosen and designed. Corresponding to the multiple cloning sites of binary plasmid p2355, XbaI and BamHI were added at the end of each strand, respectively. After being synthesized and annealed, the fragments were inserted into p2355driven by CaMV35S promotor. Identified by PCR and sequencing, the results showed that 3 vectors with double -stranded RNA-expressing construct were constructed and had potency to produce RNAi against the virus in tobacco. This could be an antiviral strategy in plant. Fig 3, Tab 1, Ref 26
- [1]S M, Elbashir,W, Lendeckel,T, Tuschl.RNA interference is mediated by 21- and 22-nucleotide RNAs.[J].2001,15(2).
- [2]Phillip D, Zamore.Ancient pathways programmed by small RNAs.[J].2002,296(5571).
- [3]Meyerowitz EM,Chuang CF.Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana.[J].2000,97(9).
- [4]K. Nowak,S. Meyer,V. K. Sharma,etc.Vectors for RNAi Technology in Poplar[J].2004,6(1).
- [5]Gregory J, Hannon,Douglas S, Conklin.RNA interference by short hairpin RNAs expressed in vertebrate cells.[J].2004,257.
- [6]Waterhouse P,Helliwell C.Constructs and methods for high-throughput gene silencing in plants.[J].2003,30(4).
- [7]A, Grishok,A E, Pasquinelli,D, Conte,etc.Genes and mechanisms related to RNA interference regulate expression of the small temporal RNAs that control C. elegans developmental timing.[J].2001,106(1).
- [8]Johansen LK,Carrington JC.Silencing on the spot. Induction and suppression of RNA silencing in theAgrobacterium-mediated transient expression system[J].2001,126(3).
- [9]Paul, Ahlquist.RNA-dependent RNA polymerases, viruses, and RNA silencing.[J].2002,296(5571).
- [10]Vanithatani R.,Fauquet CM.,Chellappan P..Short interfering RNA-mediated interference of gene expression and viral DNA accumulation in cultured plant cells[J].2003,100(16).
- [11]Helliwell CA,Waterhouse PM.Exploring plant genomes by RNA-induced gene silencing.[J].2003,4(1).
- [12]Tuschl T,Elbashir SM,Lendeckel W,etc.Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate.[J].2001,20(23).
- [13]Yuan B.,Latek R.,Hossbach M.,etc.siRNA Selection Server: an automated siRNA oligonucleotide prediction server[J].2004.
- [14]A, Fire,S, Xu,M K, Montgomery,etc.Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.[J].1998,391(6669).