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利用RT-PCR结合RACE技术克隆获得了无色书虱Ldace1的3′端序列和Ldace2的全长序列(GenBank登录号分别为:FJ647186和FJ647187)。其中Ldace2基因全长2111bp,ORF1917bp,编码638个氨基酸组成的前体蛋白,成熟蛋白的分子量为71.9kDa,理论等电点为4.70,序列比对表明该基因包括AChE家族的所有保守性功能位点;Ldace1基因仅获得了3′端1616bp的序列,编码500个氨基酸残基,序列比对发现,除缺少形成一对二硫键的半胱氨酸外,Ldace1包含了AChE家族的所有保守性功能位点。结合其它物种AChE构建的分子进化树可以看出,AChE分为4个类群:昆虫Ⅰ型AChE类群、脊椎动物AChE类群、昆虫Ⅱ型AChE类群和线虫AChE类群,其中无色书虱Ldace1和Ldace2分别属于昆虫Ⅰ型AChE类群和昆虫Ⅱ型AChE类群,且与嗜卷书虱和嗜虫书虱的AChE具有最近的进化关系。
The 3 ’end of Ldace1 and the full length of Ldace2 were cloned by RT-PCR and RACE (GenBank accession numbers: FJ647186 and FJ647187, respectively). The Ldace2 gene was 2111bp in length and contained ORF1917bp, which encoded a protein of 638 amino acids. The molecular weight of the mature protein was 71.9kDa and the theoretical isoelectric point was 4.70. Sequence alignment showed that the gene included all conserved functional sites of AChE family ; Ldace1 gene obtained only 3’1616bp sequence, encoding 500 amino acid residues, sequence alignment found that, except for the lack of a pair of disulfide bond formation of cysteine, Ldace1 contains AChE family of all conserved functions Site. According to the molecular phylogenetic tree constructed by AChE from other species, AChE is divided into four groups: the AChE group, the AChE group, the AChE group and the AChE group. Among them, Ldace1 and Ldace2 Belonging to the AChE group of insect type Ⅰ and the AChE group of insect type Ⅱ, and had the closest evolutionary relationship with AChE of the tick-boring tick and the tick-boring tick.