黑曲霉3928植酸酶phy基因的克隆及全序列分析

Cloning And sequence analysis of the phytase phya gene of aspergillus niger 3928

根据GenBank中已注册的黑曲霉基因序列设计了1对引物,通过PCR方法将黑曲霉phy A全基因进行扩增,获得了1条长约1.5 kb的特异性PCR产物,在PMD18 T载体中克隆了黑曲霉3928植酸酶phy A全基因,筛选到2个重组菌落(1#和2#),并进行了全序列测定。序列分析结果表明:2个重组质粒的测序结果完全一致;克隆的片段中含有完整的phy A基因序列,全长1 506 bp,其中含有一段长102 bp的内含子;phyA基因全序列编码467个氨基酸,信号肽位于基因的5′端,长度为19个氨基酸;碱基序列与GenBank中报道的AY513749的同源性为98.87%,编码的氨基酸序列的同源性为97.21%。

The phyA encoding phytase of Aspergillus niger 3928 was amplified by the ploymerse with the primers designed according to the sequence of the phyA in GenBank. The amplified chain reaction （PCR） fragment was clonged in PMD18 -7 Vector. The results of the sequence show ：the recoding reginin eludes the wholly gene of phyA, and the size is 1 506 bp, intron of 102 bp is in it. It encodes a peptide of 448 amino acide. Comparison with the AY513749 of GenBank,the homology of the nucleotide is as high as 98.87%, and the homogoly of the amino acide comes up to 97.21%.