摘要
目的构建与按蚊唾液腺抗疟原虫子孢子入侵相关的差异表达cDNA文库,为进一步筛选、克隆按蚊唾液腺抗疟原虫子孢子入侵差异新基因奠定基础。方法以大劣按蚊-约氏疟原虫为实验模型,分别将饱吸约氏疟原虫感染鼠血和正常鼠血后14 d的大劣按蚊作为T组和D组,采用抑制差减杂交方法和T/A克隆技术构建按蚊唾液腺抗疟原虫子孢子入侵相关cDNA文库。结果扩增差减cDNA文库获得2 670个阳性克隆,随机挑取克隆测序显示有与冈比亚按蚊未知功能基因同源的克隆。结论成功构建与按蚊唾液腺抗疟原虫子孢子入侵相关的差异表达cDNA文库。
Objective To construct a differentially expressed cDNA library of Anopheles dirus glands with anti-Plasmodium sporozoite invasion-related,lay the foundation for screening and cloning new and specific glands with anti-Plasmodium sporozoite invasion-related genes.Methods utilizing the Anopheles dirus-Plasmodium Yoelii model,the course of suppression subtractive hybridization（SSH）and T/A cloning techniques were used to construct a differentially expressed cDNA library of Anopheles dirus salivary gland,and through the analysis of EST,moleculars correlated with sporozoites invasion salivary gland would be screened.Results The amplified library contained more than 2670 positive bacteria clones.Random analysis of positive clones and the BLAST search revealed that some genes were homologous to functionally unknown Anopheles gambiae EST.Conclusion A differentially expressed glands genes subtracted cDNA library of infected An.dirus is successfully constructed.
出处
《成都医学院学报》
CAS
2011年第2期104-106,共3页
Journal of Chengdu Medical College
基金
国家自然科学基金资助项目(No.30771870
30901245)
重庆市自然科学基金资助项目(NO.2009BB321)
第三军医大学青年基金资助项目(NO.2009XQNO1)
关键词
按蚊
约氏疟原虫
抑制差减杂交
基因文库
Anopheles
Plasmodium yoeii
suppression substractive hybridiztion
gene library
