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来自嗜热栖热放线菌的木糖异构酶基因允许使用D-木糖作为选择剂有效地筛选转基因植物细胞。

The xylose isomerase gene from Thermoanaerobacterium thermosulfurogenes allows effective selection of transgenic plant cells using D-xylose as the selection agent.

作者信息

Haldrup A, Petersen S G, Okkels F T

机构信息

Danisco Biotechnology, Copenhagen K, Denmark.

出版信息

Plant Mol Biol. 1998 May;37(2):287-96. doi: 10.1023/a:1005910417789.

Abstract

The xylose isomerase gene (xylA) from Thermoanaerobacterium thermosulfurogenes (formerly Clostridium thermosulfurogenes) has been expressed in three plant species (potato, tobacco, and tomato) and transgenic plants have been selected on xylose-containing medium. The xylose isomerase gene was transferred to the target plant by Agrobacterium-mediated transformation. The xylose isomerase gene was expressed using the enhanced cauliflower mosaic virus (CaMV) 35S promoter and the omega' translation enhancer sequence from tobacco mosaic virus. Unoptimized selection studies showed that, in potato and tomato, the xylose isomerase selection was more efficient than the established kanamycin resistance selection, whereas in tobacco the opposite was observed. Efficiency may be increased by optimization. The xylose isomerase system enables the transgenic cells to utilize xylose as a carbohydrate source. It is an example of a positive selection system because transgenic cells proliferate while non-transgenic cells are starved but still survive. This contrasts to antibiotic or herbicide resistance where transgenic cells survive on a selective medium but non-transgenic cells are killed. The results give access to a new selection method which is devoid of the disadvantages of antibiotic or herbicide selection.

摘要

嗜热栖热放线菌(原嗜热硫化梭菌)的木糖异构酶基因(xylA)已在三种植物(马铃薯、烟草和番茄)中表达,并在含木糖的培养基上筛选出了转基因植株。木糖异构酶基因通过农杆菌介导的转化转移到目标植物中。木糖异构酶基因利用增强型花椰菜花叶病毒(CaMV)35S启动子和烟草花叶病毒的ω'翻译增强子序列进行表达。未优化的筛选研究表明,在马铃薯和番茄中,木糖异构酶筛选比已确立的卡那霉素抗性筛选更有效,而在烟草中则观察到相反的情况。通过优化可能会提高效率。木糖异构酶系统使转基因细胞能够利用木糖作为碳水化合物来源。它是一个正向选择系统的例子,因为转基因细胞增殖而未转基因细胞饥饿但仍存活。这与抗生素或除草剂抗性不同,在抗生素或除草剂抗性中,转基因细胞在选择培养基上存活但未转基因细胞被杀死。这些结果提供了一种新的筛选方法,该方法没有抗生素或除草剂筛选的缺点。

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