Cloning, expression and purification of pyruvate, orthophosphate dikinase regulatory proteins from Phyllostachys edulis
Pyruvate orthophosphate dikinase regulatory protein (PPDK-RP) is a key protein in C4 photosynthesis and can modified enzyme activity of PPDK.For Phyllostachys edulis,an important economic bamboo species,cloning PPDK-RP gene and studying its functions had vital theoretical value and application for bamboo photosynthesis research.Firstly PeRP1 was successfully cloned by reverse transcription polymerase chain reaction (RT-PCR),and afterward a multiple sequence alignment and phylogenetic analysis was conducted.Then for further study the crystal structure of the PeRP protein,the recombinant expression pe-SUMO vectors of PeRP were constructed and the protein was expressed in E.coli and purified by nickle beads column and size column.Results showed that the PeRP gene contained a 1.275 kb open reading frame (ORF) coding a 425 amino acid polypeptide,which contained the typical domain of UDF299 and was predicted as a number of kinase-PPPase superfamily.The multiple sequence alignment and phylogenetic analysis of PeRP revealed that Phyllostachys edulis was a typical C3 monocotyledon.Expressed soluble PeRP1 protein had three forms--polymer,dimmers,and monomers in water soultion.These provided a foundation for the structure and function of RP.
国家科技期刊平台
NSTL
万方数据
维普
