The xylose isomerase gene from Thermoanaerobacterium thermosulfurogenes allows effective selection of transgenic plant cells using D-xylose as the selection agent.
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.
Haldrup
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A
Danisco Biotechnology, Copenhagen K, Denmark.
Petersen
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SG
Okkels
F T
FT
D019747:Aldose-Ketose Isomerases / Q000235:genetics* / Q000378:metabolism; D003013:Clostridium / Q000201:enzymology / Q000235:genetics*; D018744:DNA, Plant / Q000032:analysis; D005798:Genes, Bacterial / Q000235:genetics*; D005819:Genetic Markers; D018551:Solanum lycopersicum / Q000201:enzymology / Q000235:genetics; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D010947:Plants, Toxic; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D011198:Solanum tuberosum / Q000201:enzymology / Q000235:genetics; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics; D014170:Transformation, Genetic; D014994:Xylose
D018744:DNA, Plant; D005819:Genetic Markers; D012333:RNA, Messenger; D018749:RNA, Plant; D014994:Xylose; D019747:Aldose-Ketose Isomerases; C023298:xylose isomerase