Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2008, Vol. 24 ›› Issue (10): 47-50.

Special Issue: 生物技术 棉花

• 农业生物技术科学 • Previous Articles     Next Articles

Studies on gene cloning of GhRGL from cotton and prokaryotic expression

Ming PENG   

  • Received:2008-06-10 Revised:2008-08-11 Online:2008-10-05 Published:2008-10-05

Abstract: The GA signaling pathway is repressed by the DELLA proteins. A cotton nucleotide with high sequence homology to Arabidopsis thaliana GAI (AtGAI) was identified in the NCBI GenBank database with the NCBI BLAST program, according to the sequence of the cotton nucleotide, full-length cDNAs and genomic coding sequences from upland cotton (Gossypium hirsutum) RGL (GhRGL) were obtained and characterized. Sequence comparisons between the sequence and all of DELLA proteins in Arabidopsis indicted which is a ortholog of AtRGL. And over-express it in prokaryotic systems, The expression construct pET22b carrying the GhRGL coding sequence was preformed, and transformed into E. coli BL21 used as a host. Induced by IPTG, GhRGL-His Tag fusion protein with molecular weight of 64kDa was successfully expressed in transformant. It is the first time to demonstrate recombinant expression of rape GhRGL gene in prokaryotic system, which paves the way for antibody preparation and the study of in situ expression of GhRGL.