| 从烟草悬浮细胞中分离液泡前体的方法研究 |
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| 引用本文： | 莫蓓莘,姜里文.从烟草悬浮细胞中分离液泡前体的方法研究[J].深圳大学学报(理工版),2005,22(2):109-112. |
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| 作者姓名： | 莫蓓莘 姜里文 |
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| 作者单位： | 1. 深圳大学生命科学学院,深圳,518060
2. 香港中文大学生物系,香港 |
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| 基金项目： | 香港自然科学基金资助项目 (CUHK4260) |
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| 摘 要： | 通过跟踪含有液泡前体特异标记物的组分, 利用细胞壁酶水解胞壁制造原生质体, 并用真针头挤压的方法裂解细胞, 用蔗糖密度梯度离心的方法分离液泡前体. 通过相差显微镜观察, 发现所分离的液泡前体保持完整的结构. 利用不同细胞器的标记物抗体进行免疫标记检测, 证明所纯化的液泡前体不含其他细胞器, 具有较高的纯度.
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| 关 键 词： | 液泡前体 蛋白分泌途径 蔗糖密度梯度离心 烟草悬浮细胞 纤维素酶 果胶酶 原生质体 免疫印迹法 |
| 文章编号： | 1000-2618(2005)02-0109-04 |
| 修稿时间： | 2005年1月17日 |
Isolation of prevacuolar compartments from tobacco suspension cells |
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| MO Bei-xin,JIANG Li-wen.Isolation of prevacuolar compartments from tobacco suspension cells[J].Journal of Shenzhen University(Science &engineering),2005,22(2):109-112. |
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| Authors: | MO Bei-xin JIANG Li-wen |
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| Affiliation: | MO Bei-xin~1 and JIANG Li-wen~2 1)College of Life Science Shenzhen University Shenzhen 518060 P. R. China2)Department of Biology The Chinese University of Hong Kong,Hong Kong |
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| Abstract: | Pure and intact prevacuolar(PVC) compartments were isolated from tobacco suspension cells. The technique involved enzymes to digest the cell wall, releasing cell contents, which were scavenged by syringe. The PVC was then isolated using sucrose gradient ultracentrifugation. The organization of the isolated PVC fractions remained intact as observed by differential interference contrast microscopy. Western blots using markers for different organelles proved that the isolated PVC fractions have high purity. |
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| Keywords: | prevacuolar compartment protein secretory pathway sucrose density gradient ultra-centrifugation tobacco suspension cells cellulase pectinase protoplast western blot |
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