Preparation and application of multiclonal antibody against plant cysteine proteinase

doi:10.3969/j.issn.1004-1524.2018.06.01

›› 2018, Vol. 30 ›› Issue (6): 881-885.DOI: 10.3969/j.issn.1004-1524.2018.06.01

• Crop Science • Previous Articles Next Articles

Preparation and application of multiclonal antibody against plant cysteine proteinase

PENG Qiqi¹, YANG Jian², LIAO Qiansheng^{1, *}, ZHANG Hengmu^{2, *}

1. College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China;
2. Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2018-03-28 Online:2018-06-20 Published:2018-07-02

Abstract

Abstract: Cysteine proteases (CysPs) are a family of important proteases that are involved in a wide range of plant physiological processes. To characterize such proteinase, a gene encoding CysP was amplified by RT-PCR from Nicotiana benthamiana plants and ligated into pEASYTM-T5 Zero vector. The insertion was sequenced and then sub-cloned into the prokaryotic expression plasmid pGEX-6P1, named as pGEX-NbCysP. The plasmid was transformed into Escherichia coli BL21plysS for inducible expression. The recombinant NbCysP fusion protein was purified with affinity chromatography and used for producing polyclonal antibody by immunizing rabbits. In immunoblotting assays, the polyclonal antibody could react strongly with the recombinant NbCysP protein as the presence of a single band, indicating that the antibody was specific against the proteinase. These results laid a foundation for further characterization of its function.

Key words: cysteine proteinase, prokaryotic expression, multiclonal antibodies

CLC Number:

PENG Qiqi, YANG Jian, LIAO Qiansheng, ZHANG Hengmu. Preparation and application of multiclonal antibody against plant cysteine proteinase[J]. , 2018, 30(6): 881-885.

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Preparation and application of multiclonal antibody against plant cysteine proteinase

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