枯草芽孢杆菌BJ3-2精氨酸脱羧酶基因speA的克隆与序列分析

根据GenBank中枯草芽孢杆菌168菌株的精氨酸脱羧酶基因speA序列设计特异性引物,提取Bacillus subtilis BJ3-2基因组DNA进行PCR扩增,并克隆至pGEM-T载体后测序。序列分析结果表明,Bacillus subtilis BJ3-2 speA基因ORF长为1 473 bp,可编码490个氨基酸,分子质量为53.53ku,基因登录号为KJ561348,与已报道枯草芽孢杆菌的speA基因核苷酸序列和氨基酸序列同源性均达93%以上,精氨酸脱羧酶氨基酸序列系统进化树分析,发现Bacillus subtilis BJ3-2的ADC与Bacillus subtilis 168亲缘关系最近,属于典型的III型PLP依赖型鸟氨酸/赖氨酸/精氨酸脱羧酶家族成员。speA基因序列分析及其蛋白保守结构域的分析,为有效控制水豆豉产品中生物胺含量的研究提供了理论依据。

Cloning and sequence analysis of Bacillus subtilis BJ3-2 arginine decarboxylase gene speA

Based on the arginine decarboxylase gene speA of Bacillus subtilis strain168 registered in GenBank, specific primers were designed using software DNAStar. The speA was amplified by PCR from genomic DNA of the B. subtilis B J3-2, and cloned to the pGEM-T vector for sequencing. The results of sequence analysis indicated that the open reading frame length of B. subtilis BJ3-2 speA gene was 1 473 bp, which could encode 490 amino acids with deduced molecular weight of 53.53 ku, and the GenBank accession number was No. KJ561348. The results showed that the speA gene ofB. subtilis B J3-2 was about 93% identical to the other similar nucleotide and amino acid sequences from GenBank. Phylogenetic tree indicat- ed that B. subtilis B J3-2 ADC was the nearest relative to B.subtilis strain 168, which belonged to the type III PLP-dependent ornithine decarboxylase/lysine decarboxylase/arginine decarboxylase family. The gene sequence and conserved structure domain analysis of speA would bring some new clue for further exploration of controlling the biogenic amine content of the fermented soybeans.