Journal of Pure and Applied MicrobiologyVol. 7 No. Special Edition

Structure Analysis, Prokaryotic Expression, and Subcellular Location of a New Protein Phosphatase 2C Protein from Maize

Lixia Liu

Agronomy Department, Dezhou University, Dezhou - 253 023, China. State Key Laboratory of Crop Biology, Tai’an - 271 018, China.

Received on 03 March 2013 and accepted on 14 April 2013

 

ABSTRACT

ZmPP2C (AY621066) is a protein phosphatase 2C mRNA that we cloned from maize previously. TMpred program analysis indicated this protein contains a significant transmembrane helix of about 20 amino acid residues; however SVMtm predicted it is not a transmembrane protein. STRIDE, Modeller and RasMol program analysis suggested the ZmPP2C protein is a globular protein containing seven alpha helixes, fourteen beta sheets, twenty-two turns and one 310 helix. The coding region of ZmPP2C mRNA was sub-cloned into expression vectors, pET30a-c(+), and introduced into E. coli BL21 (DE3) for expression. SDS-PAGE analysis indicated ZmPP2C was highly expressed at 37°C for 4 h with induction by 1 mM IPTG. Ultrasonic extraction experiment showed this recombinant protein was soluble in lysis buffer solution. The hexahistidine-tagged ZmPP2C fusion protein were purified and used to immunize rat for producing antibody. Protein gel blot analysis stated clearly that a specific polyclonal antibody against the protein was produced and can be used for further study about the ZmPP2C gene. Subcellular localization suggested that ZmPP2C protein was located in cell nucleus.

Keywords : Subcellular location, Prokaryotic expression, Maize, Protein phosphatase 2C, Protein gel blot.