Amplification and Expression of a Lectin Gene from <i>Agaricus bisporus</i> AS2796

LIN Yong; XIAO Dong-lai; LIU Jun-feng; TANG Xiao-luan

doi:10.19303/j.issn.1008-0384.2020.04.011

Volume 35 Issue 4

Apr. 2020

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LIN Y, XIAO D L, LIU J F, et al. Amplification and Expression of a Lectin Gene from Agaricus bisporus AS2796 [J]. Fujian Journal of Agricultural Sciences，2020，35（4）：437−442 doi: 10.19303/j.issn.1008-0384.2020.04.011

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LIN Y, XIAO D L, LIU J F, et al. Amplification and Expression of a Lectin Gene from Agaricus bisporus AS2796 [J]. Fujian Journal of Agricultural Sciences，2020，35（4）：437−442 doi: 10.19303/j.issn.1008-0384.2020.04.011

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Amplification and Expression of a Lectin Gene from Agaricus bisporus AS2796

doi: 10.19303/j.issn.1008-0384.2020.04.011

1.
Fujian Health College, Fuzhou, Fujian　350101, China
2.
Institute of Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350003, China

Received Date: 2020-01-14
Rev Recd Date: 2020-03-06
Publish Date: 2020-04-01

Abstract

Abstract

Objective A lectin gene from Agaricus bisporus AS2796 was isolated to study the functions of the agglutinin protein in mushrooms. Method Based upon the reported sequences, a primer was designed to amplify the lectin gene with the total DNA as templates. A 432 bp PCR product was purified and connected to the prokaryotic expression vector pET-28a carrying the His label, then transformed into E. coli BL21 (DE3). The IPTG-induced expression product was subsequently purified by affinity chromatography and identified by Western blot. Result The amplified lectin gene was 432 bp with correctly constructed recombinant expression plasmid, pET-lectin. The recombinant agglutinin protein was about 18 kDa and more than 95% pure after purification, Conclusion For the first time, a lectin gene was successfully isolated from A. bisporus and amplified. The agglutinin protein obtained could be used to further study its bioactivity and biosynthesis.
- Agaricus bisporus,
- lectin,
- vector construction,
- prokaryotic expression

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References(33)

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