为了获得高产植酸酶菌株,利用PCR技术,从黑曲霉WP1中克隆出2种植酸酶基因phyA和phyB,分别命名为phyA1、phyB1.phyA1长1346 bp,成熟肽序列不含内含子,共编码448个氨基酸;phyB1基因长1560 bp,基因序列含有3段内含子,共编码460个氨基酸.phyA1和phyB1序列同源性很低,只有21.12%.二者都含有1个植酸酶基因保守序列RHGXRXP;但phyA1含2个HD保守序列,phyB1只含有1个.将黑曲霉WP1植酸酶phyA1和phyB1的基因序列在国际基因库中注册,注册号分别为:DQ60711;DQ836360.
In order to get a high phytase activity strain,the phyA1 and phyB1 encoding phytase of Aspergillus niger WP1 were amplified by the polymerase chain reaction(PCR)with primers designed according to the sequences in Gene Bank.The amplified fragments were cloned and sequenced.The results showed that the coding region were 1346 bp and 1560 bp in size,there was no intron in phyA1 gene,encodes a peptide of 448 amino acids;but phyB1 includes 3 introns,and encodes a peptide of 460 amino acid residues.The area of highest homology RHGXRXP was found in the two sequences.There were two HD in phyA1 and only one in phyB1.The phyA1 and phyB1 of A.niger WP1 strain in this paper their sequences have been accessed by Gene Bank(Accession :DQ650711;DQ836360).
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