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Bulletin of Botanical Research ›› 2008, Vol. 28 ›› Issue (3): 310-314.doi: 10.7525/j.issn.1673-5102.2008.03.014

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Construction of TMV-replicase Gene-targeted Vectoer for RNA Interference

LÜFeng-Xia;GUO Zhao-Kui;YAN Pei-Qiang;WAN Xiu-Qing;PAN Yong-Ming   

  1. (1.Heilongjiang August First Land reclamation university,Daqing 163000) (2.Heilongjiang Tobacco Research Institute,Mudanjiang 157011) (3.Mudanjiang Normal university College,Mudanjiang 157012)
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-05-20 Published:2008-05-20
  • Supported by:

Abstract: The TMV-replicase gene was regarded target-directed sequence for RNA interference in this paper. The target sequence was linked to pMD18-T Simple Vector and replicated in E.coli DH 5α. after obtained by RT-PCR reaction. The pMD18-T Simple vector (which contains target sequence) was digested restrictively by double-endonuclease, after purifying the digested product was inserted into pUCCRNAi vector by the counterrepeat way. Then the counterrepeated structure of target sequence was inserted into pC2300-35s-OCS expression vector after enzyme disgested. The recombination plamid was transferred into Agrobacterium tumefaciens which only contained assistant plasmid in order to construct double-plasmid expression vector. With PCR reaction and restriction endonuclease digestion, the target gene sequence was comfirmed in engineering-bacterium.

Key words: TMV-replicase gene, RNA interference, vectoer construction

CLC Number: