The maize Knotted1 gene is an effective positive selectable marker gene for Agrobacterium-mediated tobacco transformation |
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Authors: | Keming Luo Xuelian Zheng Yongqin Chen Yuehua Xiao Degang Zhao Richard McAvoy Yan Pei Yi Li |
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Institution: | (1) Biotechnology Research Center, Southwest University, Chongqing, 400716, P.R. China;(2) Department of Biotechnology, College of Life Sciences, Hubei University, Wuhan, 430062, P.R. China;(3) Key Laboratory of Agricultural Biotechnology, Guizhou University, Guizhou, 550025, P.R. China;(4) Department of Plant Science, University of Connecticut, Storrs, CT 06269, USA |
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Abstract: | We have assessed the use of a homeobox gene knotted1 (kn1) from maize as a selectable marker gene for plant transformation. The kn1 gene under the control of cauliflower mosaic virus 35S promoter (35S::kn1) was introduced into Nicotiana tabacum cv. Xanthi via Agrobacterium-mediated transformation. Under nonselective conditions (without antibiotic selection) on a hormone-free medium (MS), a large
number of transgenic calli and shoots were obtained from explants that were infected with Agrobacterium tumefaciens LBA4404 harboring the 35S::kn1 gene. On the other hand, no calli or shoots were produced from explants that were infected with an Agrobacterium strain harboring pBI121 (nptII selection) or from uninfected controls cultured under identical conditions. Relative to kanamycin selection conferred by
nptII, the use of kn1 resulted in a 3-fold increase in transformation efficiency. The transgenic status of shoots obtained was confirmed by both
histochemical detection of GUS activity and molecular analysis. The results presented here suggest that kn1 gene could be used as an effective alternative selection marker with a potential to enhance plant transformation efficiency
in many plant species. With kn1 gene as a selection marker gene, no antibiotic-resistance or herbicide-resistance genes are needed so that potential risks
associated with the use of these traditional selection marker genes can be eliminated. |
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Keywords: | Transformation Selectable marker kn1 gene ipt gene |
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