CAJ | 학술논문

为建立一套适合茶树雌蕊总蛋白质分离的双向电泳体系，对茶树雌蕊总蛋白的提取方法、SDS-PAGE凝胶浓度、聚焦条件和IPG胶条pH范围等进行了比较优化。结果表明，TCA/丙酮-clean up kit法提取总蛋白，用17 cm pH 5~8 IPG胶条，13%SDS-聚丙烯酰胺凝胶进行分离，用考马斯亮蓝R250染色，可以获得清晰、分辨率高、重复性好的双向电泳图谱，共检测到约1200个蛋白点，主要分布在pH 5~8，相对分子量14~117 kD范围内，碱性蛋白得到有效分离，可以满足茶树雌蕊的蛋白组学分析和研究。
위건립일투괄합다수자예총단백질분리적쌍향전영체계，대다수자예총단백적제취방법、SDS-PAGE응효농도、취초조건화IPG효조pH범위등진행료비교우화。결과표명，TCA/병동-clean up kit법제취총단백，용17 cm pH 5~8 IPG효조，13%SDS-취병희선알응효진행분리，용고마사량람R250염색，가이획득청석、분변솔고、중복성호적쌍향전영도보，공검측도약1200개단백점，주요분포재pH 5~8，상대분자량14~117 kD범위내，감성단백득도유효분리，가이만족다수자예적단백조학분석화연구。
In order to establish a proper two-dimensional gel electrophoresis (2-DE) protocol for proteomic study of tea plant pistils, several protein extraction methods, density of SDS-PAGE gel, IEF procedures and pH gradient of IPG strip were compared. The results showed the optimized system includes: total proteins of sample extracted using TCA/acetone method followed with clean up, separating the proteins with 17 cm pH 5~8 IPG strips, 13%SDS-PAGE, and CBB R-250 stain method. About 1 200 spots can be detected, and these proteins mainly distributed in the pH 5~8, MW 14~117 kD range. With good separation of the basic protein, this will be helpful to proteomic research in tea plant.