摘要: |
为了研究MreB 在钝顶螺旋藻Spirulina platensis形态建成中的作用, 克隆了mreB基因并进行原核表达, 对表达的融合蛋白进行了纯化, 免疫小鼠制备了MreB的多克隆抗体, 分别用Western blot和免疫荧光技术检测不同形态藻丝体中MreB的表达和定位, 结果显示MreB表达量在螺旋形和直线形两种藻丝体中无明显差异; 免疫荧光定位显示, 荧光主要分布于细胞膜下一圈, 侧面可观察到荧光呈双股螺旋状分布。实验结果说明, MreB含量不是导致螺旋藻形态改变的因素, MreB细胞骨架蛋白参与指导细胞壁肽聚糖的合成, 所以有可能通过其双螺旋结构在藻细胞中的不对称分布影响螺旋藻的形态。 |
关键词: 螺旋形藻丝体 直线形藻丝体 细胞骨架蛋白 |
DOI:10.11759/hykx20140323003 |
分类号: |
基金项目:国家自然科学基金项目(40306024, 31301978); 湖南农田杂草防控协同创新中心资助项目 |
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Expression and Localization of MreB in Spirulina platensis |
WU Juan,JIN Chen-zhong,HU Yi-hong,HU Jun-he,ZHU Xi-wu,XU Hong
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Abstract: |
To explore the function of MreB in morphogenesis of Spirulina platensis, we cloned mreB gene into expression vector pGEX-6P-1 and expressed GST-tagged MreB in Escherichia coli. The purified MreB protein was used to immunize mice to prepare polyclonal antibodies. No obvious differential expression of MreB was detected by western blot analysis between the spiral filaments and linear forms of S. platensis. Immunofluorescence analysis showed that MreB was distributed as a ring or double helix under the cell membrane. These results showed that MreB might not directly regulate the shapes of the cell. The helical MreB cytoskeletal protein directs the synthesis of lateral peptidoglycan by providing a track, therefore, it may affect the morphogenesis of S. platensis by the asymmetric distribution. |
Key words: spiral filament linear filament Spirulina platensis cytoskeletal protein |