大劣按蚊AdPPO2、AdPPO3基因cDNA克隆与组织定位  

Molecular cloning and localization of cDNAs of AdPPO2 and AdPPO3 of Anopheles dirus

作　　者：郝宏兴[1] 邱宗文[1] 段建华[1] 况明书[1] 黄复生[1] 王英[1] 

机构地区：[1]第三军医大学基础医学部病原生物学教研室,重庆400038

出　　处：《第三军医大学学报》2004年第6期470-472,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 30 2 0 0 2 37)~~

摘　　要：目的　克隆大劣按蚊PPO基因家族新成员的cDNA部分序列 ,并对所克隆基因进行蚊体组织定位研究。方法　根据已报道的多种昆虫PPO的保守氨基酸序列设计简并引物 ,以大劣按蚊总RNA为模板进行RT PCR扩增、目的片段克隆入T载体 ,然后 ,用PCR方法筛选新的PPO克隆并测序 ;以RT PCR方法分析AdPPO2和AdPPO3在蚊血淋巴、蚊胃和唾液腺的分布情况。结果　AdPPO2和AdPPO3基因cDNA部分序列长均为 5 97bp ,编码 199个氨基酸 ;它们在血淋巴、蚊胃和唾液腺均有分布。结论　从大劣按蚊成功克隆获得 2种新的PPO基因家族成员的cDNA部分序列 。Objective To clone the partial cDNA sequences of new members of prophenoloxidase (PPO) gene family from Anopheles dirus and to localize their mRNA in the mosquitoes. Methods Degenerative primers were designed according to the conserved amino acid sequence within PPOs of insects. RNA sequence of adults of Anopheles dirus was reversely transcripted and amplified. The obtained PPO cDNA was cloned into T vector. The new AdPPOs were obtained by screening of the positive clones using PCR and sequencing. The localization of their mRNA in the hemolymph, midguts, and salivary glands was analyzed by RT PCR. Results The partial cDNA sequences of AdPPO2 and AdPPO3 were 597 bp and their deduced amino acid sequences were 199aa. Their mRNA could be found in the hemolymph, midguts, and salivary glands. Conclusion The partial cDNA sequences of the two AdPPO members from Anopheles dirus have been successfully cloned, providing the foundation for cloning their full length sequences.

关 键 词：大劣按蚊 前酚氧化酶 基因克隆 

分 类 号：Q55[生物学—生物化学] Q785