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首页> 外文期刊>Mycoscience >Cloning and functional characterization of gpd and alpha-tubulin promoters from Annulohypoxylon stygium, a companion fungus of Tremella fuciformis
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Cloning and functional characterization of gpd and alpha-tubulin promoters from Annulohypoxylon stygium, a companion fungus of Tremella fuciformis

机译：来自Annulohypoxylon Stegium的GPD和α-肾小管素启动子的克隆和功能性表征，Invella Fucifordis的伴真菌伴真菌

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摘要

Annulohypoxylon stygium is an ascomycete that helps Tremella fuciformis produce the fruiting body in wild state or artificial cultivation. Although the interaction between these two fungi is well known, the underlying molecular mechanism is limited. In this study, the 981 bp and 886 bp promoter sequences of glyceraldehyde-3-phosphate dehydrogenase (gpd) gene and alpha-tubulin gene have been cloned, respectively. Sequence analysis showed that gpd promoter contained nine CAAT boxes, and single TGACG-motif, TATA box, ABRE motif, STRE motif, MYB motif, and W box. The alpha-tubulin promoter included eight CAAT boxes, three STRE, two TATA boxes and MYB, single Box 4, CAT-box, CCAAT-box, TGA-element, and ABRE. Subsequently, we evaluated the promoters' function by constructing four vectors pGEH, pGRH, pTEH, and pTRH to drive fused enhanced green fluorescent protein and hygromycin B phosphotransferase (egfp-hph) or red fluorescent protein and hygromycin B phosphotransferase (rfp-hph) expression under the control of gpd or alpha-tubulin promoters in A. stygium. The integration of the transformed DNA into A. stygium genome was verified by PCR, Southern blot, fluorescence microscopy, and quantitative realtime PCR (qRT-PCR). All the results indicated that the two promoters could drive egfp-hph and rfp-hph expression. This result could provide help in gene functional studies by using gpd and alpha-tubulin promoters to direct gene over-expression or build dual promoter silencing systems. (C) 2019 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved.
机译：Annulohypoxylon Stegium是一种ascycete，有助于金蝶菌肌腱在野生状态或人工培养中产生果实。虽然这两个真菌之间的相互作用是众所周知的，但是潜在的分子机制是有限的。在该研究中，分别克隆了981bp和886bp甘油醛-3-磷酸脱氢酶（GPD）基因和α-微管蛋白基因的启动子序列。序列分析表明，GPD启动子含有九个Caat盒，单个TGACG-MOTIF，TATA盒，ABRE主题，StrE主题，MYB主题和W盒子。 α-小蛋白促进剂包括八个Caat盒，三个，两个塔塔盒和MYB，单箱4，猫盒，CCAAT盒，TGA元素和ABRE。随后，通过构建四个载体PGEH，PGRH，PTEH和PTRH来评估促进剂的功能，以驱动融合增强的绿色荧光蛋白和潮霉素B磷酸转移酶（EGFP-HPH）或红荧光蛋白和潮霉素B磷酸转移酶（RFP-HPH）表达在A.Stygium的GPD或α-微管蛋白启动子的控制下。通过PCR，Southern印迹，荧光显微镜和定量实时PCR（QRT-PCR）验证转化的DNA进入A. Stegium基因组的整合。所有结果表明，两个启动子可以驱动EGFP-HPH和RFP-HPH表达。该结果可以通过使用GPD和α-微管蛋白启动子直接基因过度表达或构建双启动子沉默系统来提供基因功能研究的帮助。 （c）2019日本的Mycological学会。 elsevier b.v出版。保留所有权利。

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  • 来源
    《Mycoscience》 |2020年第1期|共8页
  • 作者

    Liu Dongmei; Zhu Hanyu; Pujiana Dwi; Zheng Liesheng; Chen Liguo; Ma Aimin;

  • 作者单位

    Huazhong Agr Univ Coll Food Sci &

    Technol Wuhan 430070 Peoples R China;

    Huazhong Agr Univ Coll Food Sci &

    Technol Wuhan 430070 Peoples R China;

    Huazhong Agr Univ Coll Food Sci &

    Technol Wuhan 430070 Peoples R China;

    Huazhong Agr Univ Coll Plant Sci &

    Technol Wuhan 430070 Peoples R China;

    Huazhong Agr Univ Coll Plant Sci &

    Technol Wuhan 430070 Peoples R China;

    Huazhong Agr Univ Coll Food Sci &

    Technol Wuhan 430070 Peoples R China;

  • 收录信息
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 Q939.5;
  • 关键词

    Glyceraldehyde-3-phosphate dehydrogenase; alpha-tubulin; Promoter; Transformation;

    机译：甘油醛-3-磷酸脱氢酶;α-小管蛋白;启动子;转型;

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