摘要
目的运用SELDI(Surface Enhanced Laser Desorption/Ionization)蛋白质芯片技术检测Vero细胞感染HSV-1后蛋白质的差异表达,从而为从蛋白质水平研究HSV-1感染的致病机制奠定基础。方法常规培养Vero细胞,感染HSV-1,孵育12 h、24 h、48 h后用细胞裂解液裂解细胞。裂解上清经蛋白定量后应用IMAC3(固定金属亲和)和WCX2(弱阳离子交换)芯片,SELDI-TOF-MS技术检测细胞感染HSV-1前后蛋白质表达的差异。结果感染24 h细胞出现明显细胞病变效应(CPE),感染12 h质谱分析即可见蛋白的差异表达。共19个蛋白表达水平发生变化,其中IMAC3芯片发现差异峰9个,WCX2芯片发现差异峰10个。结论 SELDI蛋白质芯片技术检测Vero细胞感染HSV-1后蛋白质的差异表达方法简便,敏感性高,从而为从蛋白质水平研究HSV-1感染的致病机制,HSV-1与宿主细胞的相互作用,及抗感染治疗靶位的寻找奠定了一定的基础。
Objective To detect different protein expression in Vero cells infected with HSV-1 using the SELDI Protein Chip platform ,which could be used as a basis to study the patho genesis of HSV - 1 infection on protein level. Methods Vero cells were cultured in vitro, and infected with HSV - 1 in good conditions. Then the cells were harvested by cell lysis after incubation for 12,24 and 48 hours respectively. After quantification , the supernatant of the lysate was tested by IMAC3 （Immobilized mental affinity capture） and WCX2 （weak cation exchange） chip on SELDI -TOF -MS preteinchip reader. Results Apparent CPE was observed in infected ceils after incubation for 24 hours. However, different protein ex- pression was obtained by MS after 12 hours. Total 19 different expressed proteins were found, among which ,9 were captured by IMAC3 chip and 10 by WCX2 chip. Conclusion Detecting different protein expression in Vero cells infected by HSV - 1 using the SELDI Protein Chip platform was simple and sensitive;the results we got could be served as a basis to study the pathogenesis of HSV - 1 and the interaction between virus and the host . In addition , it could help the discovery of new therapy targets of HSV - 1 infection.
出处
《中国医学创新》
CAS
2010年第2期65-67,共3页
Medical Innovation of China
