| 钝顶螺旋藻表达载体p215 t-spp的构建及转化 |
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| 引用本文： | 王芬,叶霁,曹媛媛,张海生,甘旭华,唐欣昀.钝顶螺旋藻表达载体p215 t-spp的构建及转化[J].激光生物学报,2009,18(1). |
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| 作者姓名： | 王芬 叶霁 曹媛媛 张海生 甘旭华 唐欣昀 |
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| 作者单位： | 安徽农业大学生命科学学院,安徽,合肥,230036 |
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| 基金项目： | 安徽省教育厅自然科学基金 |
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| 摘 要： | 用合适的限制性内切酶消化载体p215t,胶回收获得含有gfp基因及amp基因大片段.设计引物,采用PCR扩增无菌钝顶螺旋藻A9藻株的强启动子片段;在T4连接酶的作用下进行体外连接重组,构建了带有螺旋藻启动子和gfp报告基因的新型表达载体p215t-spp.将该载体转入钝顶螺旋藻,利用报告基因的表达,在荧光显微镜下观察并记录转化藻细胞,p215t-spp质粒显著提高转化率.研究了不同PEG浓度、转化时间及冰浴处理对转化率的影响.采用1%PEG能有效促进细胞吸收DNA,获得10.5‰的转化率.实验初步证明,带有螺旋藻启动子的报告基因gfp能够在钝顶螺旋藻中顺利表达.
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| 关 键 词： | 钝顶螺旋藻 启动子 gfp基因 PEG转化 转化率 |
Construction and Transformation of Expression Vector p215t-spp of Spirulina platensis |
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| WANG Fen,YE Ji,CAO Yuan-yuan,ZHANG Hai-sheng,GAN Xu-hua,TANG Xin-yun.Construction and Transformation of Expression Vector p215t-spp of Spirulina platensis[J].ACTA Laser Biology Sinica,2009,18(1). |
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| Authors: | WANG Fen YE Ji CAO Yuan-yuan ZHANG Hai-sheng GAN Xu-hua TANG Xin-yun |
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| Affiliation: | College of Life Science;Anhui Agricultural University;Hefei 230036;Anhui;China |
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| Abstract: | The plasmid p215t was digested by the restriction enzymes and the fragment with gfp and amp gene was recovered.The promoter fragment of Spirulina A9 strain was obtained by PCR.Then the fragments above were recombined by T4 DNA ligase,and the new plasmid p215t-spp with gfp and amp gene was constructed.The new plasmid was transformed into Spirulina platensis.The evidences of fluorescence microscopy observation and electrophoretogram proved that the plasmid had been constructed successfully.Plasmid p215t-spp c... |
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| Keywords: | Spirulina platensis promoter gfp gene PEG transformation rate |
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