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Bulletin of Botanical Research ›› 2014, Vol. 34 ›› Issue (4): 492-497.doi: 10.7525/j.issn.1673-5102.2014.04.011

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

SUN Yu;WANG Jin-Lin*   

  1. 1.The Department of Ecological Engineering,Heilongjiang Vocational Institute of of Ecological Engineering,Harbin 150040;2.Graduate school,Northeast Forestry University,Harbin 150040
  • Received:1900-01-01 Revised:1900-01-01 Online:2014-07-20 Published:2014-07-20
  • Contact: WANG Jin-Lin
  • Supported by:

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-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.

Key words: rice, zinc finger domain, Prokaryotic Protein Expression and Purification

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