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首页> 外文OA文献 >Cloning of the gene for glutamate decarboxylase and its expression during conidiation in Neurospora crassa.
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Cloning of the gene for glutamate decarboxylase and its expression during conidiation in Neurospora crassa.

机译：谷氨酸脱羧酶基因的克隆及其在结节孢菌分生孢子形成过程中的表达

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摘要

Neurospora crassa glutamate decarboxylase (GAD) is produced during conidiation and stored in dormant conidia. Polyclonal antibody was generated to GAD that had been purified to homogeneity. The anti-GAD antibody was specific for N. crassa GAD and inhibited GAD activity. The level of GAD protein decreased during conidial germination, indicating that GAD was degraded during this phase of development. The anti-GAD antibody was used to isolate a cDNA clone of GAD from a lambda ZAP cDNA expression library. Escherichia coli containing a plasmid with the cDNA insert produced GAD activity. The cDNA clone contained a 2.6 kbp insert and hybridized to a 2.6 kb mRNA species from conidiating cultures of N. crassa. GAD mRNA was not present in vegetative hyphae. In conidiating cultures, GAD mRNA was first detected when conidia began to appear. The level of GAD mRNA increased as conidiation progressed. This is the first example of the cloning of an enzyme that is regulated at the level of mRNA during the asexual developmental cycle of N. crassa.
机译：分生孢子产生神经孢子谷氨酸脱羧酶（GAD），并储存在休眠分生孢子中。产生针对GAD的多克隆抗体，该抗体已纯化至同质。抗GAD抗体特异于景天猪笼草GAD，并抑制GAD活性。分生孢子萌发过程中GAD蛋白水平降低，表明GAD在该发育阶段被降解。抗GAD抗体用于从λZAP cDNA表达文库中分离GAD的cDNA克隆。含有带有cDNA插入片段的质粒的大肠杆菌产生GAD活性。该cDNA克隆含有一个2.6kbp的插入片段，并与来自N.crassa的伴生培养物的一个2.6kb的mRNA种杂交。营养菌丝中不存在GAD mRNA。在分生孢子培养物中，分生孢子开始出现时首先检测到GAD mRNA。 GAD mRNA的水平随着孕育的进行而增加。这是克隆一种酶的克隆的第一个例子，该酶在克雷萨氏菌的无性发育周期中受mRNA水平的调节。

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    Hao, R; Schmit, J C;

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  • 年度 1993
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  • 正文语种 en
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