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一种四步法提取分离白念珠菌总蛋白质方法的建立
许懿1,阎澜1,张军东2,姜远英1*
0
(1.第二军医大学药学院药理学教研室,上海 200433;2.第二军医大学长海医院皮肤科,上海 200433)
摘要:
目的:建立白念珠菌总蛋白质分步提取分离的方法。方法:利用四种溶解能力不同的溶液分步提取白念珠菌总蛋白,定量后,分别进行一维等电聚焦分离和二维聚丙烯酰胺凝胶电泳分离。结果:成功分步提取获得白念珠菌不同溶解度的蛋白,得到各步提取蛋白的二维凝胶电泳图谱。结论:四步法提取白念珠菌总蛋白质是研究白念珠菌蛋白质组的有效方法,为进一步研究白念珠菌耐药机制以及抗真菌药物作用机制奠定基础。
关键词:  白念珠菌  蛋白质组  四步法  电泳,凝胶,双向
DOI:10.3724/SP.J.1008.2007.00999
基金项目:国家自然科学基金(30630071,30672626);上海市基础研究重点项目(04JC14003)
Establishment of a “Four-step method” for extraction and separation of Candida albicans total proteins
XU Yi1,YAN Lan1,ZHANG Jun-dong2,JIANG Yuan-ying*
(1.Department of Pharmacology,School of Pharmacy,Second Military Medical University,Shanghai 200433,China; 2. Department of Dematology,Changhai Hospital,Second Military Medical University,Shanghai 200433)
Abstract:
Objective:To introduce a “Four-step method” for extraction and separation of Candida albicans total proteins.Methods: Proteins of C. albicans were extracted step-by-step with 4 kinds of solutions with different solubilities. After quantification,the protein samples were separated by isoelectric focusing electrophoresis and then by SDS-PAGE. Results: Proteins with different solubilities were successfully extracted step-by-step from C. albicans and were separated by two-dimensional gel electrophoresis. Conclusion: The “Four-step method” for extraction of C. albicans proteins is an effective approach to study C. albicans membrane proteome and lays a foundation for further investigation of the mechanisms of antifungal agents and drug resistance in C. albicans.
Key words:  Candida albicans  proteome  four-step method  electrophoresis,gel,two-dimensional