萝卜抗真菌蛋白基因Rs-AFPs在大肠杆菌中的表达及其转化番茄的研究

园艺学报 ›› 2001, Vol. 28 ›› Issue (4): 361-363.

萝卜抗真菌蛋白基因Rs-AFPs在大肠杆菌中的表达及其转化番茄的研究

邓晓东¹;费小雯²;胡新文¹

(¹ 热带作物生物技术国家重点实验室, 海口571101; 􀀁 ²中山大学生命科学学院生物工程中心, 广州 510275)

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2001-08-25 发布日期:2001-08-25

The Expression of Raphnus sativus-Antifungal Protein Gene and Its Transformation into Tomato

Deng Xiaodong¹, Fei Xiaowen², and Hu Xinwen¹

(¹National Key Biotechnology Laboratory for Tropical Crop s, CATAS , Haikou 571101; 􀀁²The College of Life Science, Zhongshan University , Guangz hou 510275)

Received:1900-01-01 Revised:1900-01-01 Online:2001-08-25 Published:2001-08-25

摘要/Abstract

摘要： 通过基因工程手段将萝卜抗真菌蛋白基因Rs-AFP_{1( 2)}插入大肠杆菌表达载体pTrxFus中并诱导表达, 其融合表达产物约22 kD, 约占总可溶蛋白的0. 45% ( 0. 5%) 。抑菌活性试验表明: 重组Rs-AFP_{1( 2)}有抑菌活性, 其中Rs􀀁AFP2 较Rs􀀁AFP1 抑菌活性强。构建了Rs-AFP₂ 基因的植物表达载体pBIAFP₂。利用农杆菌介导法将pBIAFP₂ 导入番茄中, 并诱导出完整的植株32 株。对其中28 株进行PCR 和PCR-Southern Blot 检测, 其中17 株呈阳性。对经PCR Southern Blot 检测呈阳性的植株进行Southern Blot 检测, 6 株呈阳性。

关键词: 萝卜抗真菌蛋白, 基因, 大肠杆菌, 表达, 转基因, 番茄

Abstract: Raphnus sativus-antifungal proteins ( Rs-AFPs ) are cysteine􀀁rich, antifungal proteins. The Rs-AFP₁₍₂₎ genes were inserted into the vector pTrxFus and transferred into E. coli GI724. The fusion protein from recombiant pTrxAFP_{1( 2)} was about 22 kD and taking up 0. 4 and 0. 5 percent of the total proteins respectively. Inhibition experiment showed that the expressed products had antifungal activity to Colletotrichum musae. Expressed products from pTrxAFP₂ showed much stronger inhibitory activity than that from pTrxAFP1. A plant expression vector pBIAFP₂ was constructed, which had been transferred into tomato by Agrobactrium mediated transformation. 32 plantlets had been obtained and 28 of them detected by PCR and Southern Blot. The results showed that 17 of them were positive, and the results of Southern Blot showed 6 of them were positive.

Key words: Raphnus sativus-antifungal proteins ( Rs-AFPs), Gene expression, E. coli, Transgenic, Tomato

中图分类号:

邓晓东;费小雯;胡新文. 萝卜抗真菌蛋白基因Rs-AFPs在大肠杆菌中的表达及其转化番茄的研究[J]. 园艺学报, 2001, 28(4): 361-363.

Deng Xiaodong;Fei Xiaowen;and Hu Xinwen. The Expression of Raphnus sativus-Antifungal Protein Gene and Its Transformation into Tomato[J]. ACTA HORTICULTURAE SINICA, 2001, 28(4): 361-363.

[1]	王晓晨, 聂子页, 刘先菊, 段伟, 范培格, 梁振昌, . 脱落酸对‘京香玉’葡萄果实单萜物质合成的影响[J]. 园艺学报, 2023, 50(2): 237-249.
[2]	张欣, 漆艳香, 曾凡云, 王艳玮, 谢培兰, 谢艺贤, 彭军. 香蕉枯萎病菌Dicer-like基因的功能分析[J]. 园艺学报, 2023, 50(2): 279-294.
[3]	任菲, 卢苗苗, 刘吉祥, 陈信立, 刘道凤, 眭顺照, 马婧. 蜡梅胚胎晚期丰富蛋白基因CpLEA的表达及抗性分析[J]. 园艺学报, 2023, 50(2): 359-370.
[4]	于婷婷, 李欢, 宁源生, 宋建飞, 彭璐琳, 贾竣淇, 张玮玮, 杨洪强. 苹果GRAS全基因组鉴定及其对生长素的响应分析[J]. 园艺学报, 2023, 50(2): 397-409.
[5]	翟含含, 翟宇杰, 田义, 张叶, 杨丽, 温陟良, 陈海江. 桃SAUR家族基因分析及PpSAUR5功能鉴定[J]. 园艺学报, 2023, 50(1): 1-14.
[6]	杨植, 张川疆, 杨芯芳, 董梦怡, 王振磊, 闫芬芬, 吴翠云, 王玖瑞, 刘孟军, 林敏娟. 枣与酸枣杂交后代果实遗传倾向及混合遗传分析[J]. 园艺学报, 2023, 50(1): 36-52.
[7]	崔建, 钟雄辉, 刘泽慈, 陈登辉, 李海龙, 韩睿, 乐祥庆, 康俊根, 王超. 结球甘蓝染色体片段替换系构建[J]. 园艺学报, 2023, 50(1): 65-78.
[8]	史洪丽, 李腊, 郭翠梅, 余婷婷, 简伟, 杨星勇. 番茄灰霉病生防菌株TL1的分离、鉴定及其生防能力分析[J]. 园艺学报, 2023, 50(1): 79-90.
[9]	何成勇, 赵晓丽, 许腾飞, 高德航, 李世访, 王红清. 草莓病毒1山东分离物全基因组分析[J]. 园艺学报, 2023, 50(1): 153-160.
[10]	罗海林, 袁雷, 翁华, 闫佳会, 郭青云, 王文清, 马新明. 蚕豆萎蔫病毒2号青海辣椒分离物的鉴定与全基因组序列克隆[J]. 园艺学报, 2023, 50(1): 161-169.
[11]	忽靖宇, 阙开娟, 缪田丽, 吴少政, 王田田, 张磊, 董鲜, 季鹏章, 董家红. 侵染鸢尾的番茄斑萎病毒鉴定[J]. 园艺学报, 2023, 50(1): 170-176.
[12]	赵雪艳, 王琪, 王莉, 王方圆, 王庆, 李艳. 基于比较转录组的延胡索组织差异性表达分析[J]. 园艺学报, 2023, 50(1): 177-187.
[13]	郑积荣, 王同林, 胡松申. 高品质番茄新品种‘杭杂603’[J]. 园艺学报, 2022, 49(S2): 103-104.
[14]	郑积荣, 王同林. 番茄新品种‘杭杂601’[J]. 园艺学报, 2022, 49(S2): 105-106.
[15]	郑积荣, 王同林. 樱桃番茄新品种‘杭杂503’[J]. 园艺学报, 2022, 49(S2): 107-108.