65kD热休克蛋白抗原的制备

Study of preparation of 65kD heat shock protein

[目的 ]在分析经典制备工艺及培养基配方的基础上 ,设计出简便、有效的改良法制备 6 5kD热休克蛋白。 [方法 ]将BCG(卡介苗 )在罗氏培养基上培养 ,然后转至苏通氏培养基内培养 ,4 2℃水浴休克 5 5min ,过滤除菌 ,浓缩测蛋白质量浓度。 [结果 ]经浓缩后的滤液用SDS -PAGE电泳分析显示为一条蛋白带 ,相对分子质量为 6 5kD。 [结论 ]表明方法可行。

To develop a more convenient and efficient method for preparation of 65 kD heat shock protein by analysis of classical preparation technique and culture medium composition. Bacillus Calmette-Guerin (BCG) was cultured in Robertson's cooked meat medium, then it was transferred to Sutong's medium. After shock stimulation in water bath at 42℃ for 55 minutes, the medium was filtrated to get rid of BCG, followed by concentration to determine the quantity of protein. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of concentrated filtration showed a band of 65 kD protein. [Conclusion] We established the desired preparation method.