AdPPO4基因部分序列的cDNA克隆与序列分析

Cloning and analysis of partial cDNA sequence of prophenoloxidase 4 gene of Anopheles dirus

目的 克隆大劣按蚊 (Andirus)前酚氧化酶 (prophenoloxidase ,PPO)基因部分序列。方法 根据已报道的多种昆虫以及冈比亚按蚊PPO基因编码的保守氨基酸序列设计简并引物 ,以大劣按蚊幼虫总RNA为模板进行RT PCR扩增 ,目的片段经TA克隆后测定其核苷酸序列 ,然后进行序列查询与比对。结果 从大劣按蚊幼虫中获得的PPO4基因(AdPPO4)部分序列长 5 97bp ,编码 199个氨基酸 ;AdPPO4与冈比亚按蚊PPO4基因的核酸和氨基酸序列同源性分别达 84%和 90 %。结论 从大劣按蚊幼虫中成功地克隆出了AdPPO4基因的部分序列 。

Objective To clone the partial cDNA sequence of prophenoloxidase (PPO) gene of Anopheles dirus . Methods Degenerative primers were designed according to the conserved sequence blocks within the prophenoloxidase of insects. RNA sequence of the larva of Anopheles dirus was amplified by RT PCR to get the prophenoloxidase cDNA which was then cloned into T vector and sequenced. The partial cDNA sequence of prophenoloxidase gene was analysed and compared with other prophenoloxidase gene of insects. Results The partial cDNA sequence of AdPPO4 was 597 bp, and its deduced amino acid sequence was 199aa. The cDNA sequence homology and amino acid sequence homology was 84% and 90%, respectively, as compared with the PPO4 gene of Anopheles gambiae . Conclusion The AdPPO4, with high sequence homology with the PPO4 gene of Anopheles gambiae , is successfully cloned from the larva of Anopheles dirus .