摘要
目的筛选犬小孢子菌抑制消减杂交(suppression subtractive hybridization,SSH)文库中的差异表达基因。方法应用斑点杂交和实时定量PCR技术筛选并检测差异基因;利用BLAST软件在GenBank数据库上进行同源序列比较。结果数据库比对:犬小孢子菌SSH文库中共得到13个差异表达序列,其中8个具有同源基因6个具有功能分类。实时定量PCR检测:FSH1和PQ-LRP基因在儿童头皮组织培养菌株(tester)RNA池中的表达量分别为儿童光滑皮肤培养菌株(driver1)的44.6倍和117倍;P-GAL4和NADH1基因在tester RNA池中的表达量分别为成人头皮组织培养菌株(driver2)的78.2倍和9.8倍。结论儿童头皮培养菌株中的FSH1和PQ-LRP基因较儿童光滑皮肤培养菌株中呈上调表达,P-GAL4和NADH1基因较成人头皮培养菌株中呈上调表达,提示上述基因可能是犬小孢子菌致头癣的重要基因。
Objective To screen the differentially expressed genes from suppression subtractive hybridization （SSH） library of microsporum canis. Methods Using dot-blot hybridization and real time PCR technique to screen and verify the differentially expressed genes and homology search was performed on GenBank using BLAST. Results Thirteen differentially expressed sequences were aquired from SSH library of M. canis and 8 of those had homologous genes and 6 of those had functions. Real time PCR analysis showed that the expression levels of FSH1 and PQ-LRP genes were 44.6-fold and 117-fold separately in M. canis in children＇s scalp tissue inducement （tester） than that in children＇s smooth skin （ driverl ）, while P-GAIA and NADH1 genes were 78.2-fold and 9.8-fold separately in tester than that in adult＇s scalp tissue （driver2）. Conclusion FSH1 and PQ-LRP genes showed up-regulated expression in children＇s scalp tissue culture strains than children＇s smooth skin culture strains, P-GAlA and NADH1 genes showed up-regulated expression than that in adult＇s scalp tissue culture strains, suggesting that the above-mentioned genes may be essential in tinea capitis caused by M. canis.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2009年第12期782-786,共5页
The Chinese Journal of Dermatovenereology
基金
国家自然科学基金(30640016)
