柳蚕卵黄原蛋白(Vg)cDNA3′端的克隆及序列分析

Cloning and Analysis of 3′end cDNA Sequence of Vitellogenin From Actias selene Hübner

柳蚕(Actias seleneHbner)是野生泌丝昆虫。从柳蚕雌蛹脂肪体中提取总RNA,根据已经解析出的其它泌丝昆虫的卵黄原蛋白cDNA序列设计特异性引物,对柳蚕卵黄原蛋白cDNA3′端进行RACE(rapid amplification ofcDNA ends)扩增,经克隆和测序得到了一条1 072 bp的cDNA片段,该序列与柞蚕(Antheraea pernyi)、天蚕(An-theraea yamamai)、野桑蚕(Bombyxmandarina)、家蚕(Bombyxmori)、樗蚕(Samia cynthia pryeri)、蓖麻蚕(Philosamiacynthia ricini)、樟蚕(Saturnia japonica)相应序列的同源性分别为82.5%、82.4%、67.0%、63.2%、80.3%、78.5%、81.0%。同源性分析表明,昆虫卵黄原蛋白的一级结构在进化上具有较高的保守性。

Accroding to the reported cDNA sequence of vitellogenin in other silk insects, the specific primers were designed. Then the partial cDNA of vitellogenin,1 072 bp, were obtained through 3′-RACE amplification using the total RNA from the fat body of female pupae of Actias selene Hǖbner as the template. The cloned sequence were 82.5% ,82.4% ,67.0% ,63. 2% ,80.3% ,78.5% and 81.0% identical to the counterparts of Antheraea pemyi, Antheraea yamamai, Bombyx mandarina, Bombyx mori, Samia cynthia pryeri, Philosamia cynthia riciniand Satumia japonica, respectively. The results of homologous analysis suggested that the amino acids sequence of the vitellogenin in insects were conserved in evolution.