中华蜜蜂蜂毒透明质酶基因的克隆与序列分析

摘要

The venomous hyaluronidase (Hya) gene of Chinese honey bee, Apis cerana cerana, was amplified by RT-PCR from total RNA of venom glands of the worker bees. The full length of its nucleotide is 1164 bp encoding a 387 amino acid polypeptide with predicted molecular weight of 42.6 kD. The alignment of AcHya amino acid sequence with other 6 Hyas shows that AcHya is most closely related to the Hya of European honey bee, A. mellifera ,with 91% amino acid identity. It also shares homology with Hya of Dolichovespul amaculata , Polistes annularis , Vespula vulgaris ,Lutzomia longipalpis and Homo sapiens (sperm) ,with 54% 、52% 、46% 、27% and 20% amino acid identity, respectively. A phylogenetic tree of those hyaluronidases was drawn by using GENETYX program, the conservation, the relationship between molecular structure and function of 7 hyaluronidases as above was compared and analysed.%从中华蜜蜂工蜂毒腺中快速抽提总RNA,用RT-PCR方法扩增得到该蜂毒的透明质酸酶(AcHya)基因,其核苷酸序列全长为1164 bp,可编码387个氨基酸残基,预计分子量为42.6kD.将所推导出的氨基酸序列与其它6种透明质酸酶(Hya)氨基酸序列比较,结果表明,AcHya与意大利蜜蜂Hya非常相似,具有91%的同源性,与长黄胡蜂Hya、常见黄胡蜂Hya、、马蜂Hya、吸血沙蠓唾腺Hya、人精液PH20Hya的同源性分别为54%、52%、46%、27%和20%.同时根据氨基酸序列用GENETYX程序绘制了7种Hya的分子系统树,并根据同源性比较分析了7种Hya的保守性、结构与功能的关系.