發(fā)布時間：2018-01-08 00:07

  本文關(guān)鍵詞：煙曲霉Bem46基因敲除株構(gòu)建及其功能初步研究　出處：《中國人獸共患病學(xué)報》2017年08期 　論文類型：期刊論文

  更多相關(guān)文章： Bem 煙曲霉 發(fā)芽率 信號傳導(dǎo)

【摘要】：目的構(gòu)建煙曲霉Bem46基因敲除株,初步明確Bem46基因在煙曲霉生長出芽和各種壓力傳導(dǎo)中的作用,以及對細(xì)胞壁形成的影響。方法采用生物信息學(xué)方法查找煙曲霉中Bem46基因,設(shè)計引物,并以pyrG作為篩選標(biāo)記構(gòu)建煙曲霉Bem46基因敲除株(ΔBem46)。觀察含不同壓力物質(zhì)培養(yǎng)基上對照菌株和ΔBem46徑向生長速度,觀察兩菌株發(fā)芽狀況以及在含有細(xì)胞壁抑制劑培養(yǎng)基上的生長狀況。結(jié)果通過序列比對在煙曲霉基因組中找到了Bem46基因,其編號為Afu7g04660,由1 116bp堿基組成,編碼311個氨基酸。使用原生質(zhì)體法得到煙曲霉ΔBem46并經(jīng)PCR和Southernblot確認(rèn)。經(jīng)觀察在含有山梨醇作為滲透壓力來源的培養(yǎng)基上ΔBem46生長明顯較對照菌株快。顯微鏡下觀察到在GMM液體培養(yǎng)基中,ΔBem46發(fā)芽速度慢于對照株。結(jié)論煙曲霉Bem46基因涉及由山梨醇引起的滲透壓壓力傳導(dǎo),并且該基因?qū)Υ龠M(jìn)孢子發(fā)芽可能有作用。
[Abstract]:Objective to construct the Bem46 knockout plant of Aspergillus fumigatus and to clarify the role of Bem46 gene in the growth of Aspergillus fumigatus and various pressure conduction. Methods the Bem46 gene in Aspergillus fumigatus was searched by bioinformatics and primers were designed. PyrG was used as the screening marker to construct the Bem46 knockout plant (螖 Bem46) of Aspergillus fumigatus. The radial growth rate of the control strain and 螖 Bem46 on the medium containing different pressure substances was observed. Results the Bem46 gene was found in the genome of Aspergillus fumigatus by sequence alignment. Its number is Afu7g04660, which is composed of 1116 BP base. The 螖 Bem46 of Aspergillus fumigatus was obtained by protoplast method and confirmed by PCR and Southernblot. The medium containing sorbitol as the source of osmotic pressure was observed. The growth of 螖 Bem46 was significantly faster than that of the control strain. It was observed in GMM liquid medium under microscope. Conclusion the Bem46 gene of Aspergillus fumigatus is involved in osmotic pressure conduction induced by sorbitol, and this gene may play an important role in promoting spores germination.
【作者單位】： 山西醫(yī)科大學(xué)第二醫(yī)院皮膚科;
【基金】：國家青年科學(xué)基金項目資助(No.81101233) 山西省青年科技研究基金(No.2010021036-2) 山西省衛(wèi)生計生委科研課題(No.2015043) 山西省回國留學(xué)人員科研資助項目(No.2016-052)聯(lián)合資助~~
【分類號】：R379
【正文快照】： 煙曲霉是自然界常見的一種絲狀真菌,在正常人群并不會由于經(jīng)常暴露于煙曲霉而發(fā)生感染。然而近年來隨著新技術(shù)的開展和免疫抑制劑的應(yīng)用,免疫力低下的患者往往由于暴露于煙曲霉而引起侵襲性曲霉病的發(fā)生,其發(fā)病率在臨床呈現(xiàn)明顯上升趨勢且病死率極高,因此探討煙曲霉感染的發(fā)病

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