Wang H, Wu Q, Liu S, Xie J, Ma M
College of Life Science, Sichuan University, Chengdu, 610064 China.
Wei Sheng Wu Xue Bao. 2001 Jun;41(3):310-4.
The phyA encoding phytase of Aspergillus niger N25 was amplified by the polymerase chain reaction (PCR) with primers designed according to the sequences of the phyA in GenBank. The amplified fragment was cloned and sequenced. The results show that: the coding region is 1506 bp in size, includes a 102 bp intron, and encodes a peptide of 476 amino acid residues, in which there is a signal peptide with 19 amino acids and a mature peptide of 448 amino acids. Comparison of this sequence with the phyA of the natural A. niger NRRL3135 (GenBank Accession: M94550), the most highly secreting-phytase strain, shows that the nucleotide homology is as high as 96.746%, and the amino acid homology comes up to 97.64%. The phyA of A. niger N25 strain in this paper is appropriate to be used to construct the phytase gene-engineering bacteria.
根据GenBank中植酸酶A(phyA)的序列设计引物,通过聚合酶链反应(PCR)扩增黑曲霉N25编码植酸酶的phyA基因。将扩增片段进行克隆和测序。结果表明:编码区大小为1506 bp,包含一个102 bp的内含子,编码一个由476个氨基酸残基组成的肽段,其中有一个由19个氨基酸组成的信号肽和一个由448个氨基酸组成的成熟肽。将该序列与天然黑曲霉NRRL3135(GenBank登录号:M94550,分泌植酸酶能力最强的菌株)的phyA进行比较,核苷酸同源性高达96.746%,氨基酸同源性达97.64%。本文中的黑曲霉N25菌株的phyA适合用于构建植酸酶基因工程菌。