MOLECULAR CLONING, PROKARYOTIC EXPRESSION AND PREPARATION OF ANTISERUM AGAINST LILY SYMPTOMLESS VIRUS TGB1 PROTEIN

Authors

  • Pinsan Xu School of Life Science and Biotechnology, Dalian University of Technology, Dalian, Liaoning 116024, China
  • Xinlei Wang School of Life Science and Biotechnology, Dalian University of Technology, Dalian, Liaoning 116024, China
  • Zhengyao Zhang School of Life Science and Medicine, Dalian University of Technology, Panjin, Liaoning 124221, China

DOI:

https://doi.org/10.24297/jaa.v7i1.5794

Keywords:

antiserum preparation, cloning, expression, Lily symptomless virus, TGB1 gene

Abstract

 The triple gene block gene TGB1 was amplified by RT-PCR from lily leaves infected with Lily symptomless virus and cloned into prokaryotic expression vector pET-28a(+). The recombinant vector was transformed into Escherichia coli strain BL21 (DE3). On induction with isopropyl β-D-1-thiogalactopyranoside,TGB1 protein was highly expressed and the molecular weight was 29 kDa (including a His-tag-containing fusion). After protein purification by Ni2+-NTA affinity chromatography, a polyclonal antibody against TGB1 was raised in mouse. Western blot analysis showed that the antiserum reacted specially with the TGB1 protein of LSV. ELISA and RT-PCR confirmed that the antiserum reacted specially with lily leaves infected with LSV, and can be used for a rapid test for LSV. The antibody produced in this work may be used for future immunohistochemistry and functional study of TGB1.

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References

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JOURNAL OF ADVANCES IN AGRICULTURE

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Published

2017-02-23

How to Cite

Xu, P., Wang, X., & Zhang, Z. (2017). MOLECULAR CLONING, PROKARYOTIC EXPRESSION AND PREPARATION OF ANTISERUM AGAINST LILY SYMPTOMLESS VIRUS TGB1 PROTEIN. JOURNAL OF ADVANCES IN AGRICULTURE, 7(1), 986–990. https://doi.org/10.24297/jaa.v7i1.5794

Issue

Vol. 7 No. 1 (2017)

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Articles

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