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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Zhejiang University Press
2010-11-01
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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. |
format | Article |
id | doaj-art-a2c597325aef4e88ae2e96eab642824f |
institution | Matheson Library |
issn | 1008-9209 2097-5155 |
language | English |
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 |