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首页> 外文期刊>Applied and Environmental Microbiology >Isolation and characterization of polygalacturonase genes (pecA and pecB) from Aspergillus flavus.
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Isolation and characterization of polygalacturonase genes (pecA and pecB) from Aspergillus flavus.

机译:黄曲霉中半乳糖醛酸酶基因(pecA和pecB)的分离和鉴定。

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Two genes, pecA and pecB, encoding endopolyglacturonases were cloned from a highly aggressive strain of Aspergillus flavus. The pecA gene consisted of 1,228 bp encoding a protein of 363 amino acids with a predicted molecular mass of 37.6 kDa, interrupted by two introns of 58 and 81 bp in length. Accumulation of pecA mRNA in both pectin- or glucose-grown mycelia in the highly aggressive strain matched the activity profile of a pectinase previously identified as P2c. Transformants of a weakly aggressive strain containing a functional copy of the pecA gene produced P2c in vitro, confirming that pecA encodes P2c. The coding region of pecB was determined to be 1,217 bp in length interrupted by two introns of 65 and 54 bp in length. The predicted protein of 366 amino acids had an estimated molecular mass of 38 kDa. Transcripts of this gene accumulated in mycelia grown in medium containing pectin alone, never in mycelia grown in glucose-containing medium, for both highly and weakly aggressive strains. Thus, pecB encodes the activity previously identified as P1 or P3. pecA and pecB share a high degree of sequence identity with polygalacturonase genes from Aspergillus parasiticus and Aspergillus oryzae, further establishing the close relationships between members of the A. flavus group. Conservation of intron positions in these genes also indicates that they share a common ancestor with genes encoding endopolyglacturonases of Aspergillus niger.
机译:从高侵染性黄曲霉菌株中克隆了编码内聚葡糖醛酸糖苷酶的两个基因pecA和pecB。 pecA基因由1,228 bp组成,编码363个氨基酸的蛋白质,预测分子量为37.6 kDa,被长度分别为58和81 bp的两个内含子中断。在高度侵袭性菌株中,果胶或葡萄糖生长的菌丝体中pecA mRNA的积累与以前鉴定为P2c的果胶酶的活性谱相匹配。含有pecA基因功能拷贝的弱侵染性菌株的转化体在体外产生P2c,证实pecA编码P2c。经测定,pecB的编码区长度为1,217 bp,被两个长度分别为65和54 bp的内含子打断。预测的366个氨基酸的蛋白质的估计分子量为38 kDa。该基因的转录物在仅含有果胶的培养基中生长的菌丝体中积累,而对于高和弱侵染性菌株则从未在含葡萄糖的培养基中的菌丝体中积累。因此,pecB编码先前标识为P1或P3的活动。 pecA和pecB与来自寄生曲霉和米曲霉的多半乳糖醛酸酶基因具有高度的序列同一性,进一步建立了黄曲霉组成员之间的紧密关系。这些基因中内含子位置的保守性还表明它们与编码黑曲霉内切聚乳酸糖苷酸酶的基因有共同的祖先。

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