Cloning of royal jelly AccMRJP1 gene from Apis cerana cerana and its expression in Escherichia coli

Three clones of AccMRJP1 were screened out from the sequenced brain cDNA library of Chinese honeybee according to the expression sequence tag (EST) of AccMRJP1. Through identifying with polymerase chain reaction (PCR) and sequencing, a AccMRJP1 clone containing an open reading frame (ORF) of 1 302 n...

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Main Authors: ZHANG Li-wen, DING Mei-hui, ZHANG Wei-guang, JIN Feng, SHEN Li-rong
Format: Article
Language:English
Published: Zhejiang University Press 2010-03-01
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.1008-9209.2010.02.001
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author ZHANG Li-wen
DING Mei-hui
ZHANG Wei-guang
JIN Feng
SHEN Li-rong
author_facet ZHANG Li-wen
DING Mei-hui
ZHANG Wei-guang
JIN Feng
SHEN Li-rong
author_sort ZHANG Li-wen
collection DOAJ
description Three clones of AccMRJP1 were screened out from the sequenced brain cDNA library of Chinese honeybee according to the expression sequence tag (EST) of AccMRJP1. Through identifying with polymerase chain reaction (PCR) and sequencing, a AccMRJP1 clone containing an open reading frame (ORF) of 1 302 nucleotides encoding a protein of 433 amino acids was determined. The AccMRJP1 had 99.8% similarity with the previously reported AccMRJP1 sequence (AY279539) in amino acid sequences. The AccMRJP1 was sub-cloned into the prokaryotic expression vector pGEX-4T-2 for fusion expression in Escherichia coli BL21. Analysis result of the SDS-PAGE showed that the expression product contained a specific band of protein about 76 ku in size and accumulated up to about 17.7% of the total bacterial proteins. The fusion protein was cross reactive with glutathione S-transferase (GST) polyclonal antibody and was purified through affinity chromatography, which confirmed the successful expression of GST-AccMRJP1. This work provids a technical base for the utilization of MRJP1 with biological engineering.
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publishDate 2010-03-01
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record_format Article
series 浙江大学学报. 农业与生命科学版
spelling doaj-art-4547cdca283c4f6f88d8477f9990b38c2025-08-20T03:33:22ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552010-03-013611912410.3785/j.issn.1008-9209.2010.02.00110089209Cloning of royal jelly AccMRJP1 gene from Apis cerana cerana and its expression in Escherichia coliZHANG Li-wenDING Mei-huiZHANG Wei-guangJIN FengSHEN Li-rongThree clones of AccMRJP1 were screened out from the sequenced brain cDNA library of Chinese honeybee according to the expression sequence tag (EST) of AccMRJP1. Through identifying with polymerase chain reaction (PCR) and sequencing, a AccMRJP1 clone containing an open reading frame (ORF) of 1 302 nucleotides encoding a protein of 433 amino acids was determined. The AccMRJP1 had 99.8% similarity with the previously reported AccMRJP1 sequence (AY279539) in amino acid sequences. The AccMRJP1 was sub-cloned into the prokaryotic expression vector pGEX-4T-2 for fusion expression in Escherichia coli BL21. Analysis result of the SDS-PAGE showed that the expression product contained a specific band of protein about 76 ku in size and accumulated up to about 17.7% of the total bacterial proteins. The fusion protein was cross reactive with glutathione S-transferase (GST) polyclonal antibody and was purified through affinity chromatography, which confirmed the successful expression of GST-AccMRJP1. This work provids a technical base for the utilization of MRJP1 with biological engineering.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.02.001<italic>Apis cerana cerana</italic>royal jelly<italic>AccMRJP1</italic>gene cloningprokaryotic expression
spellingShingle ZHANG Li-wen
DING Mei-hui
ZHANG Wei-guang
JIN Feng
SHEN Li-rong
Cloning of royal jelly AccMRJP1 gene from Apis cerana cerana and its expression in Escherichia coli
浙江大学学报. 农业与生命科学版
<italic>Apis cerana cerana</italic>
royal jelly
<italic>AccMRJP1</italic>
gene cloning
prokaryotic expression
title Cloning of royal jelly AccMRJP1 gene from Apis cerana cerana and its expression in Escherichia coli
title_full Cloning of royal jelly AccMRJP1 gene from Apis cerana cerana and its expression in Escherichia coli
title_fullStr Cloning of royal jelly AccMRJP1 gene from Apis cerana cerana and its expression in Escherichia coli
title_full_unstemmed Cloning of royal jelly AccMRJP1 gene from Apis cerana cerana and its expression in Escherichia coli
title_short Cloning of royal jelly AccMRJP1 gene from Apis cerana cerana and its expression in Escherichia coli
title_sort cloning of royal jelly accmrjp1 gene from apis cerana cerana and its expression in escherichia coli
topic <italic>Apis cerana cerana</italic>
royal jelly
<italic>AccMRJP1</italic>
gene cloning
prokaryotic expression
url https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.02.001
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AT zhangweiguang cloningofroyaljellyaccmrjp1genefromapisceranaceranaanditsexpressioninescherichiacoli
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