The Gene Cloning,Prokaryotic Expression and Purification of Rice Zinc Finger Protein OsRZ3

Abstract

This research is based on the NCBI's NO. AK062094,OsRZ3 gene,and uses it to design specific primer. Then through adding the cDNA's full length genome by RT-PCR,nucleotide sequencing,and compare the homology of nucleotide to show C2HC-zinc finger domain. Analyze and forecast protein molecular mass-34kD and isoelectric point is 9. 05; Arabidopsis protoplast the cytoplasm location detect indicates it is in the nucleus. Structure pGEX6P-3: : OsRZ3 fusion vector,transformation of electrical Escherichia coli BL21( DE3)bacterial strain is induced by 1 mmol·L- 1IPTG,SDS-pAGEX protein electrophoresis to show it can reach the maximum 60 kD fusion protein through 4 hours; in LB liquid culture,0. 5 mmol·L- 1IPTG is induced and express. We can get a lots of fusion protein through GST adsorption column. 1L bacterial suspension can induce and purify to concentration 1. 58 mg · mL- 1inclusion body fusion protein. The system of fusion protein pronucleus can express pGEX6P-3: : OsRZ3 fusion protein,0. 5 mmol·L- 1IPTG is induced,we get a lots of fusion protein through GST adsorption column. This is the basis of further antibody preparation and chromatin co-immunoprecipitation and the DNA combine.