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- Title
Construction of RNAi Expression Vector against Riboflavin Synthase Gene.
- Authors
Xiuyan REN; Jie QIAO; Jiangli ZHANG
- Abstract
[Objective] This study aimed to construct RNAi expression vector against riboflavin synthase (RS) gene. [Method] By using the primers designed based on RS gene coding sequence that was screened from Arabidopsis cDNA library, the 476 bp cDNA fragment of RS was amplified from pGADT7-RS recombinant plasmid, and then cloned into pUCm-T vector to obtain pUCm-RS. Two RS fragments (476 bp) were obtained through digesting pUCm-RS with restriction enzymes PstI/BamHI and PstI/XhoI, and then respectively connected into vector pBSSK-in to form pBSSK-RS-in-RS, in which the two RS fragments were inverted repeats. Finally, the transform unit RS-intron-RS, got by digesting vector pBSSK-RS-in-RS with Sac I and Kpn I, was ligated into expression vector pCAMBIA1301 to obtain the RS gene silencing vector. [Result] The restriction enzyme digestion sequencing analysis proved that the RS gene silencing vector was successfully constructed. [Conclusion] This study may provide a basic: material for further studies on the bio-function of RS gene and the mechanism of signal transduction induced by HpaGXoo in plant.
- Subjects
RNA interference; RIBOFLAVIN synthase; ARABIDOPSIS; ANTISENSE DNA; CLONING
- Publication
Agricultural Biotechnology (2164-4993), 2012, Vol 1, Issue 2, p43
- ISSN
2164-4993
- Publication type
Article