<em>FpgMBV1</em>-<em>P4</em>基因的原核表达及多克隆抗体制备

doi:10.3969/j.issn.1674-7968.2022.11.017

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Abstract Fusarium pseudograminearum megabirnavirus 1 (FpgMBV1) is a mycovirus found in the hypovirulence Fusarium pseudograminearum strain FC136-2A. Four proteins were encoded in FpgMBV1. Among them, FpgMBV1-P4 composed of 268 amino acids with unknown function. Specific primers were designed and used in reverse transcription PCR (RT-PCR) to amplify the FpgMBV1-P4 gene from the hyphae of the strain FC136-2A in this study. Then a prokaryotic expression vector pGEX4T-FpgMBV1-P4 was constructed and transformed into Escherichia coli BL21(DE3) strain, and the recombinant protein was induced at 25 ℃ with 0.5 mmol/L isopropyl β-D-thiogalactoside (IPTG). Results showed that FpgMBV1-P4 in length of 807 bp was successfully amplified and the fusion protein with a molecular weight of about 60 kD was highly expressed . The purified fusion protein was used to immunize Zealand white rabbit (Lepus sinensis) to obtain antiserum. Indirect ELISA results showed the titer of this antiserum reached 1∶160 000. And the antiserum was used to specifically detect the FpgMBV1-P4 protein at about 36 kD from samples of the hyphae of FC136-2A. These results laid a foundation for the structural and functional analysis of FpgMBV1-P4, which would be meaningful in deciphering the hypovirulent mechanism of FpgMBV1.

Key words： Mycovirus Fusarium pseudograminearum megabirnavirus 1 (FpgMBV1) P4 protein Prokaryotic expression Polyclonal antibody

Received: 24 November 2021

ZTFLH:

S432.1

Corresponding Authors: * Corresponding authors, zhangxiaoting921@163.com; daij666@126.com

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	LIU Dong-Wei
	SONG Jia-Qing
	PAN Xin
	YAN Shu-Wei
	LI Ke
	FAN Pei
	GAO Fei
	ZHANG Xiao-Ting
	DAI Jun-Li
	LI Hong-Lian

Cite this article:

LIU Dong-Wei,SONG Jia-Qing,PAN Xin, et al. Prokaryotic Expression of FpgMBV1-P4 Gene and Preparation of Its Polyclonal Antibodies[J]. 农业生物技术学报, 2022, 30(11): 2246-2254.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2022.11.017 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2022/V30/I11/2246

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