摘要
目的利用DNA条形码信息研制DNA芯片,建立快速高效的医学媒介蚊类鉴定方法。方法以蚊科5属15种,共30个样本为研究材料,获取其DNA条形码信息(COⅠ基因片段序列),采用基因块motif技术,搜索15个样本的同源保守基因序列,制作虚拟DNA条形码芯片。虚拟电子杂交30个样本,应用软件Bio-Edit计算属、种及种内个体间的同一性,应用phylip3.66软件以最大简约法分别对杂交结果和原序列进行聚类分析比较。结果选择78个DNA片段制作一张虚拟DNA条形码芯片,计算属间的平均同一性为0.75,同一属内种间的平均同一性为0.84,种内个体间的平均同一性为0.98,聚类分析结果一致。结论研制的虚拟DNA芯片能够鉴定本研究中的15种蚊虫,利用DNA条形码信息,设计DNA芯片在理论上可行。
Objective To validate the feasibility of the high effective quick method establishment for the medical vector mosquito identification by DNA chip. Methods Thirty CO I gene partial sequences of fifteen mosquitoes which belong to five genus were analyzed. The homologous gene sequences of fifteen sequences were searched by motif and a DNA barcode chip in silico was generated. The thirty CO I gene partial sequences were hybridized in silico, and the identities of different genus, different species in the same genus and different samples of one specie were analyzed by Bio-Edit. The results of hybridized in silico and DNA sequence were clustered and compared using maximum parsimony method by phylip 3.66. Results A DNA barcode chip in silico with seventy-eight DNA fragment was generated. The average identity of different genus was 0.75, and the average identity of different species in the same genus was 0.84, and the average identity of different samples in the same specie was 0.98. The cluster from the results of hybridized in silico was consistent with the cluster from DNA sequence. Conclusion The DNA barcode chip can identify the fifteen mosquitoes in silico, and integrating the DNA barcode and DNA chip is feasible.
出处
《中国媒介生物学及控制杂志》
CAS
CSCD
北大核心
2008年第2期99-103,共5页
Chinese Journal of Vector Biology and Control
基金
国家自然科学基金(30570250)
国家质检总局科技计划项目(2006IK189)
