摘要
目的采用抑制性消减杂交(SSH)方法筛选与克隆果蝇七氟烷敏感性相关的基因,以进一步研究吸入麻醉药的作用机制。方法以对七氟烷敏感和耐药的果蝇品系为研究模型,以野生型黑腹果蝇作为对照,应用SSH方法筛选与吸入麻醉药七氟烷敏感性相关的候选基因。结果筛选到的差异片段中,在敏感品系中高调的有77个,低调的有56个;在耐药品系中高调的有58个,低调的有52个;在敏感品系中高调而耐药品系中低调的有4个。经过与果蝇基因库比对,确定了其中大部分基因的功能,发现这些基因均非位于Y染色体上;与吸入麻醉药敏感性相关的基因大部分位于第2号染色体上。结论本实验结果与先前有的学者用遗传学方法定位所得结果基本一致,七氟烷麻醉相关基因定位于2号染色体上,为常染色体显性遗传,并且果蝇对吸入麻醉药的敏感性在雌性与雄性间无差异,为进一步研究吸入麻醉药的作用机制提供了新的思路和方法。
Objective To explore the molecular mechanisms of pharmacological action of inhaled anesthetics by screening and cloning the sevoflurane sensitivity genes in three different strains of Drosophila melanogaster. Methods Suppression subtractive hybridization （SSH） was applied to detect mRNA levers of sevoflurane sensitivity genes in the sevoflurane-sensitive strain, the sevoflurane-resistant strain and the wild type strain of Drosophila melanogaster; totally 243 differential fragments （cDNA） were identified by SSH. Results Compared to wild type strain, there were 77 upregulated and 56 downregulated cDNAs in sevoflurane-sensitive strain, 58 upregulated and 52 downregulated cDNAs in sevoflurane-resistant strain, four cDNAs were upregulated in sevofluranesensitive strain while downregulated in sevoflurane-resistant strain. All sevoflurane-sensitive cNDAs were located in x-chromosome （Chr. 2）. Conclusion Genotyping profiles are associated with sevoflurane sensitivity in the three different strains of Drosophila melanogaster,which may cast lights on the pharmacological mechanisms of inhalation anesthetics.
出处
《上海医学》
CAS
CSCD
北大核心
2009年第1期65-68,共4页
Shanghai Medical Journal
基金
国家自然科学基金资助项目(30170908)
关键词
抑制性消减杂交
果蝇
七氟烷
敏感
耐药
基因
Suppression subtractive hybridization
Drosophila melanogaster
Sevoflurane, Sensitive, Resistant, Gene
