Construction of plastid transient expression vector of GFP gene and its expression in E. Coli

Abstract

Green fluorescent protein (GFP) has recently been developed as a monitoring marker for gene expression and protein localization in living organisms In this study,a plastid transient expression vector, pJGFP, was constructed by replacing the coding region of psbA gene with the GFP cDNA gene.The GFP was highly expressed in Escherichia coli DH5α under the control of the promoter of psbA gene,and the protein was readily detected by fluorescent spectrophotometer. The results show that GFP expressed in E.coli is capable of producing a strong green fluorescence (wavelength 509 nm), when excited by blue light (wavelength 395 nm). The GFP expressed in E.coli could also be seen in SDS-PAGE In this paper, we have also discussed the potential applicationof the plastid transient expression vector pJGFP as monitoring marker for plastid transformation