通过构巢曲霉中的位点特异性整合转化对β-微管蛋白基因进行鉴定和功能分析。
Identification and functional analysis of beta-tubulin genes by site specific integrative transformation in Aspergillus nidulans.
作者信息
May G S, Gambino J, Weatherbee J A, Morris N R
出版信息
J Cell Biol. 1985 Sep;101(3):712-9. doi: 10.1083/jcb.101.3.712.
Abstract
We have cloned two different beta-tubulin sequences from the filamentous fungus Aspergillus nidulans. Each was used in the construction of transforming plasmids that carry the pyr4 gene of Neurospora crassa. We used these plasmids to transform a pyrG-strain of Aspergillus to uridine prototrophy. Both plasmids were shown to integrate site specifically into the homologous chromosomal sequences. We then used transformant strains in genetic crosses to demonstrate that one of the cloned beta-tubulin sequences was the benA beta-tubulin gene, which codes for the beta 1-and beta 2-tubulins. The other cloned beta-tubulin sequence was shown to be the structural gene for beta 3-tubulin by gene disruption and to participate in conidial development. This is the first report of a gene disruption by site specific, integrative recombination in Aspergillus nidulans.
摘要
我们从丝状真菌构巢曲霉中克隆了两个不同的β-微管蛋白序列。每一个序列都被用于构建携带粗糙脉孢菌pyr4基因的转化质粒。我们用这些质粒将构巢曲霉的一个pyrG菌株转化为尿苷原养型。结果表明，这两个质粒都能位点特异性地整合到同源染色体序列中。然后，我们在遗传杂交中使用转化菌株，证明其中一个克隆的β-微管蛋白序列是benAβ-微管蛋白基因，它编码β1和β2微管蛋白。通过基因破坏实验表明，另一个克隆的β-微管蛋白序列是β3微管蛋白的结构基因，并参与分生孢子的发育。这是关于构巢曲霉中通过位点特异性整合重组进行基因破坏的首次报道。
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