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黄曲霉β-微管蛋白基因的分离、序列分析及其作为选择标记的应用。

Isolation and sequence analysis of a beta-tubulin gene from Aspergillus flavus and its use as a selectable marker.

作者信息

Seip E R, Woloshuk C P, Payne G A, Curtis S E

机构信息

Department of Plant Pathology, North Carolina State University, Raleigh 27695-7616.

出版信息

Appl Environ Microbiol. 1990 Dec;56(12):3686-92. doi: 10.1128/aem.56.12.3686-3692.1990.

DOI:10.1128/aem.56.12.3686-3692.1990
PMID:2128007
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC185052/
Abstract

An altered beta-tubulin gene that confers resistance to benomyl [whose active ingredient is 2-(methoxycarbonylamino)benzimidazole (MBC)] was isolated from a DNA library of Aspergillus flavus and used as a selectable marker for transformation. The beta-tubulin gene was cloned into a plasmid vector containing the pyr-4 gene of Neurospora crassa, and transformants were selected either for uracil prototrophy or MBC resistance. Transformants selected for uracil prototrophy were of three phenotypic classes: sensitive, intermediate, and resistant to MBC. Transforming DNA appeared to integrate at several sites in the genome, with the more resistant phenotypes having more copies of the altered beta-tubulin gene than the sensitive and intermediate phenotypes. Transformants were also selected on medium containing MBC. The average frequency of transformation (1 to 3 transformants per micrograms of transforming DNA) was lower than that obtained by selection for uracil prototrophy, presumably because of failure to select transformants that contained few copies of the altered beta-tubulin gene. The sequence of the beta-tubulin gene was determined and compared with the published sequence of the benA gene of A. nidulans; the beta-tubulin gene was found to be highly conserved between the two Aspergillus species. Notable differences were that the beta-tubulin gene of A. flavus lacks intron 6 present in benA and has an additional leucine at position 148. This is the first gene sequence reported from an aflatoxin-producing fungus and adds to the growing body of knowledge of the beta-tubulin genes and their use as selectable markers for transformation of filamentous fungi.

摘要

从黄曲霉的DNA文库中分离出一个赋予对苯菌灵（其活性成分是2-(甲氧基羰基氨基)苯并咪唑（MBC））抗性的β-微管蛋白基因变体，并将其用作转化的选择标记。将β-微管蛋白基因克隆到含有粗糙脉孢菌pyr-4基因的质粒载体中，通过尿嘧啶原养型或MBC抗性筛选转化体。筛选出的尿嘧啶原养型转化体有三种表型类别：对MBC敏感、中间型和抗性。转化DNA似乎整合到基因组的几个位点，抗性越强的表型比敏感和中间型表型含有更多拷贝的改变后的β-微管蛋白基因。也在含有MBC的培养基上筛选转化体。转化的平均频率（每微克转化DNA有1至3个转化体）低于通过尿嘧啶原养型筛选获得的频率，推测是因为未能筛选到含有少量改变后的β-微管蛋白基因拷贝的转化体。测定了β-微管蛋白基因的序列，并与已发表的构巢曲霉benA基因序列进行比较；发现这两个曲霉物种之间的β-微管蛋白基因高度保守。显著的差异是黄曲霉的β-微管蛋白基因缺乏benA中存在的内含子6，并且在第148位有一个额外的亮氨酸。这是首次报道的产黄曲霉毒素真菌的基因序列，增加了关于β-微管蛋白基因及其作为丝状真菌转化选择标记的应用的知识。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4d2/185052/a1f98ba37782/aem00093-0070-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4d2/185052/62c0db44a8e0/aem00093-0069-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4d2/185052/9af96361eb98/aem00093-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4d2/185052/66d3d86a4c2c/aem00093-0070-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4d2/185052/a1f98ba37782/aem00093-0070-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4d2/185052/62c0db44a8e0/aem00093-0069-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4d2/185052/9af96361eb98/aem00093-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4d2/185052/66d3d86a4c2c/aem00093-0070-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4d2/185052/a1f98ba37782/aem00093-0070-c.jpg

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本文引用的文献

1
Efficient cloning of genes of Neurospora crassa.高效克隆粗糙脉孢菌基因。
Proc Natl Acad Sci U S A. 1986 Jul;83(13):4869-73. doi: 10.1073/pnas.83.13.4869.
2
A cosmid for selecting genes by complementation in Aspergillus nidulans: Selection of the developmentally regulated yA locus.通过互补作用在构巢曲霉中选择基因的 cosmid：发育调节的 yA 基因座的选择。
Proc Natl Acad Sci U S A. 1985 Feb;82(3):834-8. doi: 10.1073/pnas.82.3.834.
3
Rapid isolation of high molecular weight plant DNA.高分子量植物DNA的快速分离
aflJ基因的特性研究，该基因是将途径中间体转化为黄曲霉毒素所必需的。
Appl Environ Microbiol. 1998 Oct;64(10):3713-7. doi: 10.1128/AEM.64.10.3713-3717.1998.
4
ord1, an oxidoreductase gene responsible for conversion of O-methylsterigmatocystin to aflatoxin in Aspergillus flavus.ord1，一种负责在黄曲霉中将O-甲基柄曲霉素转化为黄曲霉毒素的氧化还原酶基因。
Appl Environ Microbiol. 1997 May;63(5):1661-6. doi: 10.1128/aem.63.5.1661-1666.1997.
5
Analysis of mechanisms regulating expression of the ver-1 gene, involved in aflatoxin biosynthesis.参与黄曲霉毒素生物合成的ver-1基因表达调控机制分析。
Appl Environ Microbiol. 1997 Mar;63(3):1058-65. doi: 10.1128/aem.63.3.1058-1065.1997.
6
Identification of aflatoxin biosynthesis genes by genetic complementation in an Aspergillus flavus mutant lacking the aflatoxin gene cluster.通过在缺乏黄曲霉毒素基因簇的黄曲霉突变体中进行遗传互补来鉴定黄曲霉毒素生物合成基因。
Appl Environ Microbiol. 1996 Oct;62(10):3567-71. doi: 10.1128/aem.62.10.3567-3571.1996.
7
Molecular analysis of the Septoria nodorum beta-tubulin gene and characterization of a benomyl-resistance mutation.小麦颖枯病菌β-微管蛋白基因的分子分析及苯菌灵抗性突变的鉴定
Mol Gen Genet. 1993 Feb;237(1-2):58-64. doi: 10.1007/BF00282784.
8
Cloning of the afl-2 gene involved in aflatoxin biosynthesis from Aspergillus flavus.从黄曲霉中克隆参与黄曲霉毒素生物合成的afl-2基因。
Appl Environ Microbiol. 1993 Jan;59(1):156-62. doi: 10.1128/aem.59.1.156-162.1993.
9
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Mol Gen Genet. 1993 Jul;240(1):73-80. doi: 10.1007/BF00276886.
10
The alcohol dehydrogenase gene adh1 is induced in Aspergillus flavus grown on medium conducive to aflatoxin biosynthesis.酒精脱氢酶基因adh1在生长于有利于黄曲霉毒素生物合成的培养基上的黄曲霉中被诱导。
Appl Environ Microbiol. 1994 Feb;60(2):670-6. doi: 10.1128/aem.60.2.670-676.1994.
Nucleic Acids Res. 1980 Oct 10;8(19):4321-5. doi: 10.1093/nar/8.19.4321.
4
A catalogue of splice junction sequences.剪接连接序列目录。
Nucleic Acids Res. 1982 Jan 22;10(2):459-72. doi: 10.1093/nar/10.2.459.
5
Codon selection in yeast.酵母中的密码子选择
J Biol Chem. 1982 Mar 25;257(6):3026-31.
6
Precursor recognition by kinetic pulse-labeling in a toxigenic aflatoxin B1-producing strain of Aspergillus.在产毒黄曲霉毒素B1的黄曲霉菌株中通过动力学脉冲标记进行前体识别。
Appl Environ Microbiol. 1981 Jul;42(1):168-73. doi: 10.1128/aem.42.1.168-173.1981.
7
Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs.真核生物mRNA翻译起始位点上游序列的汇编与分析。
Nucleic Acids Res. 1984 Jan 25;12(2):857-72. doi: 10.1093/nar/12.2.857.
8
Genetics of Aspergillus flavus: linkage of aflatoxin mutants.黄曲霉的遗传学：黄曲霉毒素突变体的连锁关系
Can J Microbiol. 1984 Jan;30(1):68-73. doi: 10.1139/m84-012.
9
Point mutations identify the conserved, intron-contained TACTAAC box as an essential splicing signal sequence in yeast.点突变确定了保守的、内含于内含子中的TACTAAC盒是酵母中一个必需的剪接信号序列。
Cell. 1984 Mar;36(3):645-53. doi: 10.1016/0092-8674(84)90344-1.
10
Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis.利用寡脱氧核苷酸定向诱变构建改良的M13载体。
Gene. 1983 Dec;26(1):101-6. doi: 10.1016/0378-1119(83)90040-9.