CAJ | 학술논문

为建立副猪嗜血杆菌蛋白质组学双向电泳方法，从裂解液配方、样品前处理、上样量以及聚焦时间对副猪嗜血杆菌临床分离株进行双向电泳体系的优化。通过超声裂解（7 mol·L－1尿素裂解液）离心法提取全蛋白，结合2-D clean up 试剂盒纯化样品，用银染方法确定了24 cm 胶条上样量为500μg，一向等电聚焦电压达到60000 Vh，可以获得重复性好、分辨率高的双向电泳图谱。
위건립부저기혈간균단백질조학쌍향전영방법，종렬해액배방、양품전처리、상양량이급취초시간대부저기혈간균림상분리주진행쌍향전영체계적우화。통과초성렬해（7 mol·L－1뇨소렬해액）리심법제취전단백，결합2-D clean up 시제합순화양품，용은염방법학정료24 cm 효조상양량위500μg，일향등전취초전압체도60000 Vh，가이획득중복성호、분변솔고적쌍향전영도보。
This study aimed to establish an efficient 2-DE proteomic method of H .parasuis .The 2-DE reaction suitable for the determination was established and optimized through the formulation of lysis buffer,modification on sample preparation,adjustment on sampling,and time needed for isoelectric focusing determination.Two clinical isolates of H .parasuis were used inthe comparative analysis.The results showed that high-resolution,high-repeatability 2-DE pages could be obtained with 500 μg protein samples extracted by the ultrasound-lysis (7 mol· L-1 ultra lysis buffer)-centrifugation method and purified by a 2-D clean up kit.They were separated by isoelectric focusing on the first dimension reaching 60 000 Vh,and visualized by silver-staining.