银杏小孢子蛋白提取及双向电泳技术优化  

Microspore Protein Extraction from Ginkgo biloba and Optimization of Two-dimensional Electrophoresis Techniques

作　　者：王亚茹[1] 李云[1] 孙宇涵[1] 王谦[2] 

机构地区：[1]北京林业大学林木育种国家工程实验室,林木花卉遗传育种教育部重点实验室,国家林业局树木花卉育种与生物工程重点开放实验室,生物科学与技术学院,北京100083 [2]北京林业大学总务与产业管理处环境科,北京100083

出　　处：《西南林学院学报》2010年第3期45-49,共5页Journal of Southwest Forestry College

基　　金：国家自然科学基金项目(30771747)资助;农业部公益性行业科研专项(200803034)资助

摘　　要：对银杏小孢子蛋白提取方法、上样量等进行试验。结果表明:在蛋白提取方面,TCA/丙酮法最佳,该方法提取的总蛋白得率最高,所获蛋白在2-DE图谱获得的点数最多、最清晰;饱和酚法次之;Tris-HCl法最差。上样量方面,蛋白上样量为1 500μg时得到的2-DE图片,蛋白点清晰,重叠现象少,图谱质量最佳;蛋白上样量为2 100μg时2-DE图谱的多蛋白点相互重叠,横竖条纹严重;上样蛋白为900μg时,2-DE图谱上蛋白点较少,丰度较弱。A methodology experiment was conducted to extract the microspore protein from Ginkgo biloba,together with sample preparation,loading dosage of the samples and gel electrophoresis procedure.The results showed that the best method to extract the microspore protein from Ginkgo biloba was TCA/acetone with the highest protein yield,the most spots in protein map,and the highest resolution of protein,and the most clear background could be obtained simultaneously.The TCA/acetone method was followed by the extraction method with saturated phenol,and the Tris-HCl extraction method.The 1 500 μg of protein loading dosage could obtain clear 2-DE map with more protein spots,better separation effect and the best spectrum.The 2 100 μg of protein loading dosage obtained map with more protein overlaps and fringes.Whereas the 900 μg protein loading dosage obtained map with less protein spots.

关 键 词：银杏 双向电泳 蛋白提取 上样量 

分 类 号：S792.950.4[农业科学—林木遗传育种]