Expression of the Glycine-ich RNA-binding Protein of Tobacco in E.coli

doi:Q786

China Biotechnology

2010, Vol. 30

Issue (08): 27-30 DOI: Q786

Expression of the Glycine-ich RNA-binding Protein of Tobacco in E.coli

LU Xiu-ping¹,CHEN Xue-jun¹,LIU Yong¹,TANG Qi-hui²,CHEN Yu-bao³,LI Wen-zheng¹

1.The Research Institute of Tobacco Science of Yunnan Province,Yuxi 653100,China
2.Beijing Biokit Science and Technology Limited Company，Beijing 100094,China
3.SinoGreen Institute for BioEconomy，Beijing 102206,China

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Abstract

Objective To clone, express the Glycine-rich RNA-binding protein of tobacco. Methods Amplified the full length NtRGP-1a and NtRGP-3 gene of tobacco, then ligated the gene into pGEX4T-1.Expression vectors pGEX4T-1/NtRGP-1a, pGEX4T-1/ NtRGP-3 were constructed and transformed into E．coli rosetta for expression induced by IPTG. Recombinant proteins were purified through GSTrap 4B affinity chromatography. Results The recombinant gene can be overexpressed in E.coli. SDS-PAGE showed that the molecular weight of the expressed product were the same as expected. The purity of the protein is greater than 95%. Conclusion the NtRGP-1a and NtRGP-3 protein of tobacco has been successful cloned and expressed, which could be useful for developing monoclonal antibody and the research of stress resistance.

Key words： Tobacco Glycine-rich RNA-binding protein Clone and express

Received: 13 April 2010 Published: 25 August 2010

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	LEI Xiu-Ping
	CHEN Hua-Jun
	LIU Yong
	TANG Qi-Hui
	CHEN Yu-Bao
	LI Wen-Zheng

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LEI Xiu-Ping, CHEN Hua-Jun, LIU Yong, TANG Qi-Hui, CHEN Yu-Bao, LI Wen-Zheng. Expression of the Glycine-ich RNA-binding Protein of Tobacco in E.coli. China Biotechnology, 2010, 30(08): 27-30.

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https://manu60.magtech.com.cn/biotech/Q786 OR https://manu60.magtech.com.cn/biotech/Y2010/V30/I08/27

［1］ Chin Y D,Grabowski P J,Sharp P A,et al.Heterogeneous nuclear ribonucleoproteins: role in RNA splicing. Science,1986,231:15341539.
［2］ Mattaj I D.Cap trimethylation of U snRNA is cytoplasmic and dependent on U snRNP protein binding.Cell,1986,46:905911.
［3］ Hall T M.Poly(A) tail synthesis and regulation :recent structural insights.Curr Opin Struct Biol,2002,12:8288.
［4］ Nomata T,Kabeya Y,Sato N.Cloning and characterization of glycinerich RNAbinding protein cDNAs in the Moss Physcomitrella patens. Plant Cell Physio,2004,45:4856.
［5］ Hanano S,Sugita M,Sugiura M.Isolation of a novel RNAbinding protein and its association with a large ribonucleoprotein particle present in the nucleoplasm of tobacco cells. Plant Mol Biol,1996,31:5768.
［6］ Van Nocker S,Vierstra R D.Two cDNA from Arabidopsis thaliana encode putative RNAbinding proteins containing glycinerich domains. Plant Mol Biol,1993,21:695699.
［7］ Carpenter C D,Krepes J A,Simon A E.Genes encoding glycinerich Arabidopsis thaliana proteins with RNA binging motifs are influenced by cold treatment and an endogenous circadian rhythm. Plant Physiol,1994,104:10151025.
［8］ Heintzen C,Naterm Apel K, Staiger D.AtGRP7, a nuclear RNAbinding protein as a component of a circadianregulation negative feedback loop in Arabidopsis thaliana. Proc Natl Acad Sci USA,1997,94:85188520.
［9］ Pereira H G, Tumova B ,Law V G. Animal influenza A virus.World Health Organization,1965,32:855860.

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