膠孢炭疽菌CgRGS2基因的克隆及生物學(xué)功能
發(fā)布時(shí)間:2018-04-17 09:47
本文選題:膠孢炭疽菌 + G蛋白信號(hào)調(diào)控因子; 參考:《微生物學(xué)報(bào)》2017年01期
【摘要】:【目的】G蛋白信號(hào)調(diào)控因子(Regulators of G-protein signaling,RGS)是G蛋白的一類負(fù)調(diào)控因子,在植物病原菌生長(zhǎng)發(fā)育及致病過(guò)程中發(fā)揮著重要的作用,然而目前還未有關(guān)于膠孢炭疽菌RGS蛋白生物學(xué)功能的研究。本試驗(yàn)的目的是克隆膠孢炭疽菌的一個(gè)RGS基因CgRGS2,并分析其生物學(xué)功能!痉椒ā坷肞CR技術(shù)擴(kuò)增CgRGS2的基因并進(jìn)行生物信息學(xué)分析,利用同源重組的方法獲得CgRGS2基因的敲除突變體,并在突變體的基礎(chǔ)上獲得互補(bǔ)株,通過(guò)表型分析確定該基因的生物學(xué)功能!窘Y(jié)果】通過(guò)PCR擴(kuò)增獲得了CgRGS2的基因,其編碼一個(gè)574個(gè)氨基酸的蛋白,在N末端含有一個(gè)RGS功能域。該基因敲除突變體同野生型相比,表現(xiàn)為營(yíng)養(yǎng)生長(zhǎng)緩慢,氣生菌絲濃密,分生孢子產(chǎn)量降低且孢子呈多端萌發(fā),對(duì)氧化壓力及SDS敏感,致病性減弱等!窘Y(jié)論】CgRGS2蛋白參與調(diào)控膠孢炭疽菌的營(yíng)養(yǎng)生長(zhǎng),分生孢子產(chǎn)量及萌發(fā),氧化應(yīng)激反應(yīng)及細(xì)胞壁完整性,對(duì)其致病性也具有一定的影響。
[Abstract]:[objective] G protein signal regulators of G-protein signaling (RGS) is a negative regulator of G protein, which plays an important role in the growth, development and pathogenicity of plant pathogens.However, there is no study on the biological function of RGS protein of Bacillus anthracis.The purpose of this study was to clone CgRGS2, a RGS gene of Bacillus anthracis, and to analyze its biological function. [methods] CgRGS2 gene was amplified by PCR and bioinformatics was performed.The knockout mutants of CgRGS2 gene were obtained by homologous recombination, and complementary strains were obtained on the basis of mutants. The biological function of the gene was determined by phenotypic analysis. [results] the gene of CgRGS2 was amplified by PCR.It encodes a 574 amino acid protein and contains a RGS functional domain at the N terminal.Compared with wild type, the knockout mutant showed slow vegetative growth, dense hyphal density, decreased conidial yield and multi-terminal germination of spores, and was sensitive to oxidative pressure and SDS.[conclusion] CgRGS2 protein is involved in the regulation of vegetative growth, conidial yield and germination, oxidative stress response and cell wall integrity of anthrax, and has a certain effect on its pathogenicity.
【作者單位】: 海南大學(xué)環(huán)境與植物保護(hù)學(xué)院;
【基金】:國(guó)家自然科學(xué)基金(31560045) 海南省自然科學(xué)基金(20153132)~~
【分類號(hào)】:S432.44
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