| 以潮霉素抗性为选择标记的毛壳霉原生质体转化 |
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| 引用本文： | 陈振明,郭泽建,林福呈,李德葆. 以潮霉素抗性为选择标记的毛壳霉原生质体转化[J]. 浙江大学学报(农业与生命科学版), 2001, 27(1): 19-22 |
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| 作者姓名： | 陈振明 郭泽建 林福呈 李德葆 |
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| 作者单位： | 浙江大学生物技术研究所，浙江 杭州 310029 |
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| 基金项目： | 国家自然科学基金资助项目(39870514) |
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| 摘 要： | 利用PEG方法，将含有潮霉素抗性标记的质粒pCB1003转化毛壳霉原生质体,并在高于毛壳霉敏感的潮霉素浓度下筛选转化子。转化率达1～2个转化子/μg质粒DNA。以潮霉素抗性的毛壳霉转化子的基因组DNA为模板进行PCR扩增及其产物的Southern杂交。结果表明，潮霉素抗性基因已整合入毛壳霉染色体。稳定性试验表明，所获得的转化子可通过有丝分裂稳定传代。
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| 关 键 词： | 毛壳霉 潮霉素抗性 转化 |
| 文章编号： | 1008-9209(2001)01-0019-04 |
Transformation ofChaetomium globosum with hygromycin B resistance plasmids pCB1003 |
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| CHEN Zhen ming,GUO Ze jian,LIN Fu cheng,LI De bao. Transformation ofChaetomium globosum with hygromycin B resistance plasmids pCB1003[J]. Journal of Zhejiang University(Agriculture & Life Sciences), 2001, 27(1): 19-22 |
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| Authors: | CHEN Zhen ming GUO Ze jian LIN Fu cheng LI De bao |
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| Abstract: | Genetic transformation has been achieved by incubatingprotoplasts from Chaetomium globosum with plasmid PCB1003. The efficiency was 1-2 transformants/μg vector DNA. Protoplasts were produced by treating 14-hour old mycelium of Chaetomium globosum with 10 mg/mL lysozyme. Chaetomium globosum growth was inhibited completely when the medium contained 150 μg/mL hygromycin B. Integration of hph gene was verified by PCR and PCR product hybridization analysis. There was no evidence of mitotic of hygromycin B resistance in the transformants. |
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| Keywords: | Chaetomium globosum transformation hygromycin B resistance |
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