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首页> 外文学位 >Protein subcellular targeting in Trichoderma aggressivum f. aggressivum.
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Protein subcellular targeting in Trichoderma aggressivum f. aggressivum.

机译：侵袭性木霉f。侵略性的

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摘要

Trichoderma aggressivum f. aggressivum is a filamentous soil fungus. Green mold disease of commercial mushrooms caused by this species in North America has resulted in millions of dollars in lost revenue within the mushroom growing industry. Research on the molecular level of T. aggressivum have just begun with the goal of understanding the functions of each gene and protein, and their expression control. Protein targeting has not been well studied in this species yet. Therefore, the intent of this study was to test the protein localization and production levels in T. aggressivum with green fluorescent protein (GFP) with an intron and tagged with either nuclear localization signal (NLS) or an endoplasmic reticulum retention signal (KDEL). Two GFP constructs (with and without the intron) were used as controls in this study.;All four constructs were successfully transferred into T. aggressivum and all modified strains showed similar growth characteristics as the wild type non-transformed isolate. GFP expression was detected from all modified T. aggressivum with confocal microscopy and the expression was similar in all four strains. The intron tested in this study had no or very minor effects as GFP expression was similar with or without it. The GFP signal increased over a 5 day period for all transformants, while the GFP to total protein ratio decreased over the same period for all transformants. The GFP-KDEL transformant showed similar protein expression level and localization as did the control transformant lacking the KDEL retention signal. The GFP-NLS transformant similarly failed to localize GFP into nucleus as fluorescence with this strain was virtually identical to the GFP transformant lacking the NLS. Thus, future research is required to find effective localization signals for T. aggressivum.
机译：侵略性木霉f。侵略性是丝状土壤真菌。在北美，由该物种引起的商品蘑菇的绿色霉菌病已导致蘑菇种植业的数百万美元的收入损失。刚草的分子水平研究刚刚开始，其目的是了解每种基因和蛋白质的功能及其表达控制。蛋白质靶向尚未在该物种中进行深入研究。因此，本研究的目的是用带有内含子的绿色荧光蛋白（GFP）并用核定位信号（NLS）或内质网保留信号（KDEL）标记，来测试侵略性小麦中的蛋白质定位和生产水平。在该研究中使用两个GFP构建体（具有和不具有内含子）作为对照。将所有四个构建体成功地转移到侵略性山毛榉中，并且所有修饰的菌株显示出与野生型未转化分离株相似的生长特性。用共聚焦显微镜从所有改良的侵略性T.aggressivum中检测到GFP表达，并且在所有四个菌株中表达相似。在这项研究中测试的内含子没有或只有很小的影响，因为有或没有GFP的表达都相似。对于所有转化体，GFP信号在5天的时间内增加，而对于所有转化体，GFP与总蛋白的比例在同一时期降低。 GFP-KDEL转化子显示出相似的蛋白质表达水平和定位，而缺少KDEL保留信号的对照转化子也是如此。 GFP-NLS转化子同样未能将GFP定位到细胞核中，因为该菌株的荧光实际上与缺少NLS的GFP转化子相同。因此，需要进行进一步的研究以找到针对侵略性山毛榉的有效定位信号。

著录项

  • 作者

    Wang, Tao.;

  • 作者单位

    Brock University (Canada).;

  • 授予单位 Brock University (Canada).;
  • 学科 Biology Molecular.
  • 学位 M.Sc.
  • 年度 2010
  • 页码 90 p.
  • 总页数 90
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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