蚊AFLP技术体系的建立  

Establishment of Mosquito AFLP System

作　　者：胡红霞[1] 孙立新[2] 叶松[1] 陆军[1] 朱国强[2] 杨庆贵[2] 

机构地区：[1]安徽理工大学医学院,安徽淮南232001 [2]江苏出入境检验检疫局

出　　处：《中国国境卫生检疫杂志》2012年第1期38-42,共5页Chinese Journal of Frontier Health and Quarantine

基　　金：国家质检总局科研基金项目(2009IK220)

摘　　要：目的探讨蚊扩增片段长度多态性(Amplified Fragment Length Polymorphism,AFLP)的各种影响因素,建立并优化蚊AFLP反应体系。方法以淡色库蚊成蚊为研究材料,对AFLP反应中的关键步骤,即多氧核糖核酸(Deoxyribonucleic acid,DNA)提取、酶切及连接反应、预扩增和选择性扩增体系进行探索。结果酶切及连接反应5h完成,将产物稀释10倍进行预扩增得到清晰的弥散带;预扩增产物稀释10倍及调整选扩循环数,可获得带型丰富且重复性好的结果。结论蚊AFLP反应体系的建立为进一步分子生物学研究提供了基础。Objective To establish the Amplified Fragment Length Polymorphism（AFLP） system of mosquito,and to optimize the key factors.Methods Culex pipiens pallens was the objective of study,the critical protocol of AFLP system, including the extraction of template DNA,restriction endonuclease digestion and ligation of adapters,preamplification and selective amplification,were explored.Results The reaction system of digestion and ligation was ended completely with 5 h,the ligation products with lμl of 10 times-diluted were used as templates for preamplification and then obtained clearly diffusible band.Meanwhile,1μl of 10 times-diluted preamplification products for selective amplification and modified the number of cycles,the reaction products derived from AFLP reaction system can produce clear,complete and detachable bands.Conclusion The AFLP system is established successfully,which provided foundation for further molecular study and gene cloning of mosquito.

关 键 词：蚊 扩增片段长度多态性 反应体系 多氧核糖核酸 

分 类 号：R187.5[医药卫生—流行病学]