%0 Journal Article
%A SUN Yu
%A WANG Jin-Lin*
%T The Gene Cloning,Prokaryotic Expression and Purification of Rice Zinc Finger Protein OsRZ3
%D 2014
%R 10.7525/j.issn.1673-5102.2014.04.011
%J Bulletin of Botanical Research
%P 492-497
%V 34
%N 4
%X 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-34 kD 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-1 IPTG ,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-1 IPTG 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-1 inclusion body fusion protein. The system of fusion protein pronucleus can express pGEX6P-3::OsRZ3 fusion protein,0.5 mmol·L-1 IPTG 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.
%U https://bbr.nefu.edu.cn/EN/10.7525/j.issn.1673-5102.2014.04.011