中国野桑蚕滞育激素基因的克隆与序列分析

Cloning and Sequence Analysis of the Gene Encoding the Precursor Protein of Diapause Hormone of Bombyx mandarina China

根据家蚕DH PBAN基因序列设计特异引物 ,以中国野桑蚕基因组DNA为模板进行PCR扩增 ,获得了1 5 85kb的目的片段 ,将其克隆进pMD18 T载体 ,测序和同源性比较结果表明 :该片段由 3个外显子、2个内含子组成 ,2个内含子均为典型的GT AG结构 :该片段编码中国野桑蚕滞育激素 (DH)及DH引导肽 ;中国野桑蚕DH及引导肽与家蚕及日本野桑蚕的DH及引导肽氨基酸序列完全相同 ,其基因cDNA序列的同源性分别为 99 3%、97 2 %。研究结果进一步显示了中国野桑蚕与家蚕之间的近缘关系。

The primers are designed according to DNA sequence of the gene encoding the precursor protein of DH PBAN of Bombyx mori to amplify the genome DNA of Chinese wild silkworm ( Bombyx mandarina China), and an aimed 1.585 kb fragment is obtained. It is cloned into pMDT 18 vector, and sequenced, the result showed that it includes 3 exons and 2 introns, which are typical of GT AG structure, and encodes DH and its leading neuropeptide. Its homologies on amino acid and nucleic acid between Bombyx mandaria China and Bombyx mori are 100%,98%, and those between Bombyx mandarina China and Bombyx mandarina Japan are 97%,97% respectively. This research further reveals the close relationship between the Bombyx mandarina China and Bombyx mori.