›› 2012, Vol. 24 ›› Issue (5): 0-763.

• 论文 •    

PolyA trapping by using terminatorless RFP reporter gene in rice

LI Jun;QYU Shao-hong*   

  1. Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2012-09-24 Published:2012-09-24

Abstract: Gene trapping method has been widely used in plant functional genomic research. PolyA trapping vector pLJ19 which contains GFP and RFP reporter genes was constructed, GFP reporter gene was used for detecting transformation events and RFP reporter gene was used for PolyA trapping. Calli both with GFP and RFP fluorescence were selected and T-DNA insertion sites were analysed by Genome walking PCR. The plasmid pLJ19 was introduced into calli of Oryza sativa L. ssp. Japonica cv. Nipponbare and then 17 transgenic calli both with GFP and RFP. FST of T-DNA insertion sites indicated that 6 T-DNA were inserted into rice genome and 5 of 6 them were inserted around rice gene 3’ ends. These results indicated that the plasmid pLJ19 was available in PolyA trapping.

Key words: gene trapping, terminator, red fluorescent protein (RFP), rice