Expression of Acc-royalisin from royal jelly of Apis cerana cerana in Escherichia coli and preparation of its polyclonal antibody

The coding region of pre-pro-Acc-royalisin was amplified by PCR from cDNA library of the Chinese honeybee, Apis cerana cerana head, and was cloned into the vector pGEX-4T-2 for expression in Escherichia coli BL21. The expressed fusion protein, glutathione S-transferase (GST)- pre-pro-Acc-royalisin o...

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Main Authors: DING Mei-hui, JIN Feng, SHEN Li-rong, CHEN Zheng-xian
Format: Article
Language:English
Published: Zhejiang University Press 2010-11-01
Series:浙江大学学报. 农业与生命科学版
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Online Access:https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.06.003
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author DING Mei-hui
JIN Feng
SHEN Li-rong
CHEN Zheng-xian
author_facet DING Mei-hui
JIN Feng
SHEN Li-rong
CHEN Zheng-xian
author_sort DING Mei-hui
collection DOAJ
description The coding region of pre-pro-Acc-royalisin was amplified by PCR from cDNA library of the Chinese honeybee, Apis cerana cerana head, and was cloned into the vector pGEX-4T-2 for expression in Escherichia coli BL21. The expressed fusion protein, glutathione S-transferase (GST)- pre-pro-Acc-royalisin of 36 ku was obtained, which was cross-reacted with GST antibody accounting for up to 16.3% of bacterial protein. With the expressed products retrieved from the SDS-PAGE gels as antigen to immunize New Zealand white rabbits, the polyclonal antibody was prepared. With the purified recombinant GST-pre-pro-Acc-royalisin fusion protein as antigen, the high titers of the antibody was shown with ELISA analysis. The specificity of the antibody against the same antigens was then confirmed by Western blot. This study provides a new tool for the detection of antimicrobial of royal jelly, biological product quality of royalisin and resistance of honeybee.
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publishDate 2010-11-01
publisher Zhejiang University Press
record_format Article
series 浙江大学学报. 农业与生命科学版
spelling doaj-art-a2c597325aef4e88ae2e96eab642824f2025-08-01T05:35:23ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552010-11-013660961410.3785/j.issn.1008-9209.2010.06.00310089209Expression of Acc-royalisin from royal jelly of Apis cerana cerana in Escherichia coli and preparation of its polyclonal antibodyDING Mei-huiJIN FengSHEN Li-rongCHEN Zheng-xianThe coding region of pre-pro-Acc-royalisin was amplified by PCR from cDNA library of the Chinese honeybee, Apis cerana cerana head, and was cloned into the vector pGEX-4T-2 for expression in Escherichia coli BL21. The expressed fusion protein, glutathione S-transferase (GST)- pre-pro-Acc-royalisin of 36 ku was obtained, which was cross-reacted with GST antibody accounting for up to 16.3% of bacterial protein. With the expressed products retrieved from the SDS-PAGE gels as antigen to immunize New Zealand white rabbits, the polyclonal antibody was prepared. With the purified recombinant GST-pre-pro-Acc-royalisin fusion protein as antigen, the high titers of the antibody was shown with ELISA analysis. The specificity of the antibody against the same antigens was then confirmed by Western blot. This study provides a new tool for the detection of antimicrobial of royal jelly, biological product quality of royalisin and resistance of honeybee.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.06.003<italic>Apis cerana cerana</italic>royal jelly<italic>Acc-royalisin</italic>prokaryotic expressionpolyclonal antibody
spellingShingle DING Mei-hui
JIN Feng
SHEN Li-rong
CHEN Zheng-xian
Expression of Acc-royalisin from royal jelly of Apis cerana cerana in Escherichia coli and preparation of its polyclonal antibody
浙江大学学报. 农业与生命科学版
<italic>Apis cerana cerana</italic>
royal jelly
<italic>Acc-royalisin</italic>
prokaryotic expression
polyclonal antibody
title Expression of Acc-royalisin from royal jelly of Apis cerana cerana in Escherichia coli and preparation of its polyclonal antibody
title_full Expression of Acc-royalisin from royal jelly of Apis cerana cerana in Escherichia coli and preparation of its polyclonal antibody
title_fullStr Expression of Acc-royalisin from royal jelly of Apis cerana cerana in Escherichia coli and preparation of its polyclonal antibody
title_full_unstemmed Expression of Acc-royalisin from royal jelly of Apis cerana cerana in Escherichia coli and preparation of its polyclonal antibody
title_short Expression of Acc-royalisin from royal jelly of Apis cerana cerana in Escherichia coli and preparation of its polyclonal antibody
title_sort expression of acc royalisin from royal jelly of apis cerana cerana in escherichia coli and preparation of its polyclonal antibody
topic <italic>Apis cerana cerana</italic>
royal jelly
<italic>Acc-royalisin</italic>
prokaryotic expression
polyclonal antibody
url https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.06.003
work_keys_str_mv AT dingmeihui expressionofaccroyalisinfromroyaljellyofapisceranaceranainescherichiacoliandpreparationofitspolyclonalantibody
AT jinfeng expressionofaccroyalisinfromroyaljellyofapisceranaceranainescherichiacoliandpreparationofitspolyclonalantibody
AT shenlirong expressionofaccroyalisinfromroyaljellyofapisceranaceranainescherichiacoliandpreparationofitspolyclonalantibody
AT chenzhengxian expressionofaccroyalisinfromroyaljellyofapisceranaceranainescherichiacoliandpreparationofitspolyclonalantibody