CAJ | 학술논문

目的建立人胆汁蛋白质组双向荧光差异凝胶电泳分析流程用于胆汁比较蛋白质组学研究.方法实验组与对照组样本各6例,分别取自于胆管癌及胆总管结石病例.样本经纯化处理及定量后,分别标记不同的荧光染料后进行双向电泳与差异分析.结果两组样本均获得高分辨力的双向电泳图谱;各标记蛋白质点的荧光强度与蛋白表达量呈线性关系;实验组与对照组间共筛选出55个差异表达蛋白,其表达量两组间相差1.5倍以上,t检验有统计学意义(P＜0.05).结论建立了基于双向荧光差异凝胶电泳基础上的胆汁比较蛋白质组学的分析流程.
목적 건립인담즙단백질조쌍향형광차이응효전영분석류정용우담즙비교단백질조학연구.방법 실험조여대조조양본각6례,분별취자우담관암급담총관결석병례.양본경순화처리급정량후,분별표기불동적형광염료후진행쌍향전영여차이분석.결과 량조양본균획득고분변력적쌍향전영도보;각표기단백질점적형광강도여단백표체량정선성관계;실험조여대조조간공사선출55개차이표체단백,기표체량량조간상차1.5배이상,t검험유통계학의의(P＜0.05).결론 건립료기우쌍향형광차이응효전영기출상적담즙비교단백질조학적분석류정.
Objective To establish a two-dimensional differential in-gel electrophorisis process for comparative proteomie analysis of human bile,Methotis Bile fluid was obtained from 12 patients with obstructive iaundice(each 6 cases of cholangioeareinoraa and choledocholith).The unfractionated bile fluid was performed sonication and ultracentrifugation to ramove the cellular debris and Nucleic Acid as a crude separation.Commercial kits Cleanaseite and mierocon were adopted to remove the abundant lipid and salt.After quantify by BCA Kit,concentrated and purified samples were labeled with three different CyDyes including one internal standard,and separated with 2-D DIGE in triplicate experiments.Biological variations of the level of protein expression were analyzed with DeCyder software and evaluated for reproducibility and statistical significance.Resuits Common gel artifacts can be reduced by above mentioned samples treatment method and high resolution biliary 2-D DIGE maps can be obtained;the fluorescence intensity of separated protein spots have a line relationship with the level of their protein expression:55 matched protein spots with statistical variance of two groups were detected(t-test,P＜0.05).Conclusion An ideal sample preparation strategy and optimized 2-D DIGE analysis process for human bile were established.