MOLECULAR CLONING, PROKARYOTIC EXPRESSION AND PREPARATION OF ANTISERUM AGAINST LILY SYMPTOMLESS VIRUS TGB1 PROTEIN
Journal Title: JOURNAL OF ADVANCES IN AGRICULTURE - Year 2017, Vol 7, Issue 1
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.
Authors and Affiliations
Pinsan Xu, Xinlei Wang, Zhengyao Zhang
Keywords
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- EP ID EP652415
- DOI 10.24297/jaa.v7i1.5794
- Views 166
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