| 毕赤酵母Mut+和Muts重组子表达美洲商陆抗病毒蛋白基因的比较 |
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| 引用本文： | 周倩,王锡锋,李莉,周广和,高必达.毕赤酵母Mut+和Muts重组子表达美洲商陆抗病毒蛋白基因的比较[J].植物病理学报,2003,33(5):425-428. |
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| 作者姓名： | 周倩 王锡锋 李莉 周广和 高必达 |
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| 作者单位： | 1 中国农业科学院植物保护研究所, 北京 100094;2 湖南农业大学植物保护学院, 长沙 410128 |
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| 基金项目： | 国家烟草专卖局重点课题(1102002010);国家科技攻关项目(2001BA509B04)资助 |
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| 摘 要： | 将N末端信号肽和C末端毒性区域缺失突变的美洲商陆抗病毒蛋白(pokeweed antiviral protein,PAP)基因表达载体pPIC9KP酶切线性化后,通过电击转化整合到巴氏毕赤酵母(Pichia pastoris) GS115菌株细胞中,PCR筛选出表型为利用甲醇快速型(Mut+)的重组子。在相同培养条件下比较Mut+重组子和利用甲醇缓慢型(Muts)重组子在表达缺失突变型PAP方面的异同。结果表明,诱导培养48 h后,Mut+重组子表达产物在SDS-PAGE胶上可见清晰目的带,而Muts重组子培养72 h才能见到微弱的目的带。抗病毒活性实验表明Mut+重组子和Muts重组子的表达产物均对TMV病毒侵染有明显的抑制作用,它们的抗病毒活性没有显著的差异。
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| 关 键 词： | 美洲商陆抗病毒蛋白 毕赤酵母 表达 Mut+重组子 Muts重组子 |
| 文章编号： | 0412-0914(2003)05-0425-04 |
| 修稿时间： | 2003年1月18日 |
The expression of pokeweed antiviral protein gene in Pichia pastoris Mut+ and Muts recombinants |
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| ZHOU Qian ,WANG Xi-feng ,LI Li ,ZHOU Guang-he ,GAO Bi-da.The expression of pokeweed antiviral protein gene in Pichia pastoris Mut+ and Muts recombinants[J].Acta Phytopathologica Sinica,2003,33(5):425-428. |
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| Authors: | ZHOU Qian WANG Xi-feng LI Li ZHOU Guang-he GAO Bi-da |
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| Institution: | 1 Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China;2 College of Plant Protection, Hunan Agricultural University, Changsha 410128, China |
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| Abstract: | The expression vector pPIC9K-P including the truncated mutant pokeweed antiviral protein (PAP) gene was li- nearized and then transformed electrically into Pichia pastoris GS115. The Mut + recombinant was selected by PCR and the expression of pokeweed antiviral protein was compared with Mut s recombinant in P. pastoris under the same condition. SDS-PAGE results showed that there was a clear target band in Mut + recombinant culture supernatant after 48 hours culture but only a faint band in Mut s recombinant culture supernatant after 72 hours culture. Activity tests revealed that two culture supernatants could inhibit plant virus infection with high efficiency and there was no significant difference between them. |
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| Keywords: | pokeweed antiviral protein PAP Pichia pastoris expression Mut + recombinant Mut s recombinant |
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