[Cloning and sequence analysis of the phytase phyA gene of Aspergillus niger N25]

Abstract

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.