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Cloning and characterization of the aldA gene of Aspergillus nidulans 总被引:13,自引:0,他引:13
M Pickett D I Gwynne F P Buxton R Elliott R W Davies R A Lockington C Scazzocchio H M Sealy-Lewis 《Gene》1987,51(2-3):217-226
We have cloned and sequenced the aldA (encoding aldehyde dehydrogenase) gene of Aspergillus nidulans. The gene contains two introns which are similar in size and structure to other fungal introns. The amino acid sequence of aldehyde dehydrogenase (497 residues) shows a significant level of homology with analogous sequences in other organisms. Comparison of the primary structure of the active sites of the mammalian cytosolic and mitochondrial enzymes shows that the Aspergillus enzyme closely resembles the mammalian mitochondrial enzyme. Analysis of the 5' non-coding region of the aldA gene shows a TATA-like sequence located 90 bp upstream from the initiation codon. Two messenger-RNA start points are located 36 and 42 bp upstream from the start codon. 相似文献
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Ribosomal -1 frameshifting during decoding of Bacillus subtilis cdd occurs at the sequence CGA AAG
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During translation of the Bacillus subtilis cdd gene, encoding cytidine deaminase (CDA), a ribosomal -1 frameshift occurs near the stop codon, resulting in a CDA subunit extended by 13 amino acids. The frequency of the frameshift is approximately 16%, and it occurs both when the cdd gene is expressed from a multicopy plasmid in Escherichia coli and when it is expressed from the chromosomal copy in B. subtilis. As a result, heterotetrameric forms of the enzyme are formed in vivo along with the dominant homotetrameric species. The different forms have approximately the same specific activity. The cdd gene was cloned in pUC19 such that the lacZ' gene of the vector followed the cdd gene in the -1 reading frame immediately after the cdd stop codon. By using site-directed mutagenesis of the cdd-lacZ' fusion, it was shown that frameshifting occurred at the sequence CGA AAG, 9 bp upstream of the in-frame cdd stop codon, and that it was stimulated by a Shine-Dalgarno-like sequence located 14 bp upstream of the shift site. The possible function of this frameshift in gene expression is discussed. 相似文献
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Cloning and selective overexpression of an alkaline protease-encoding gene from Aspergillus oryzae. 总被引:3,自引:0,他引:3
The gene alpA encoding Aspergillus oryzae alkaline protease (ALP) was isolated from a genomic library of an industrial strain used in Thailand by using oligodeoxyribonucleotide probes based on the published cDNA sequence [Tatsumi et al., Agric. Biol. Chem. 52 (1988) 1887-1888]. The entire nucleotide sequence of the genomic clone obtained was determined. By comparison with the published cDNA sequence, it was found that ALP is encoded by four exons of 314, 445, 89 and 351 bp. Three introns, which interrupt the coding sequence, are 50, 59 and 56 bp in length. The gene contains a typical TATA box 103 bp upstream from the start codon, and a consensus polyadenylation signal, AATAAA, 189 bp from the stop codon. The alpA gene, introduced into a protease deficient strain (A. oryzae U1638) by cotransformation, directed the secretion of enzymatically active ALP into the culture medium. Cotransformants of the high-level ALP-producing strain U212 containing multiple copies of the alpA gene were able to secrete up to five times more ALP than the parental strain. 相似文献
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运用同源克隆的方法设计简并引物,通过3′和5′RACE技术,从石蒜科植物朱顶兰(Amaryllis vittata Ait)总RNA中克隆了编码此凝集素(AVA)的全长cDNA序列.该基因全长686 bp,起始密码子位于第41~43 bp,终止密码子位于515~517 bp处,开放阅读框长474 bp,编码158个氨基酸,包含信号肽序列、成熟蛋白序列和C-末端剪切序列的前体蛋白.成熟蛋白由109个氨基酸残基组成,分子量为11.9kD.成熟蛋白在氨基酸水平上与雪花莲凝集素、水仙凝集素、石蒜凝集素和君子兰凝集素分别有73.4%、85.3%、80.7%和83.5%的同源性;朱顶兰凝集素的分子模式显示其与雪花莲凝集素有极其相似的三维结构;在Blocks数据库中检索AVA蛋白氨基酸序列的结构域,发现有3个凝集素功能结构域,并具有3个典型的甘露糖专一结合位点盒(QDNY). 相似文献
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运用同源克隆的方法设计简并引物,通过3′和5′RACE技术,从石蒜科植物朱顶兰(Amaryllis vittata Ait)总RNA中克隆了编码此凝集素(AVA)的全长cDNA序列。该基因全长686 bp,起始密码子位于第41~43 bp,终止密码子位于515~517bp处,开放阅读框长474 bp,编码158个氨基酸,包含信号肽序列、成熟蛋白序列和C-末端剪切序列的前体蛋白。成熟蛋白由109个氨基酸残基组成,分子量为11.9kD。成熟蛋白在氨基酸水平上与雪花莲凝集素、水仙凝集素、石蒜凝集素和君子兰凝集素分别有73.4%、85.3%、80.7%和83.5%的同源性;朱顶兰凝集素的分子模式显示其与雪花莲凝集素有极其相似的三维结构;在Blocks数据库中检索AVA蛋白氨基酸序列的结构域,发现有3个凝集素功能结构域,并具有3个典型的甘露糖专一结合位点盒(QDNY)。 相似文献
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Genetic transformation of Monascus purpureus DSM1379 总被引:4,自引:0,他引:4
Monascus purpureus was transformed into hygromycin B resistance with hygromycin B phosphotransferase (hph) fused to Aspergillus nidulans trpC or a putative Monascus purpureus gpd1 promoter by electroporation. Among five strains, only M. purpureus DSM1397 was a competent recipient. Normal growth and sporulation on media containing up to 500 mg hygromycin B l–1 occurred up to five generations. Upon transformation of the strain with the green fluorescent protein gene (sgfp) as a model gene and hph as a selection marker, characteristic green fluorescence was observed under fluoromicroscopy indicating successful transformation. 相似文献
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