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| Developing transgenic maize (Zea mays L.) with insect resistance and glyphosate tolerance by fusion gene transformation |
| SUN He, LANG Zhi-hong, LU Wei, ZHANG Jie, HE Kang-lai , ZHU Li, LIN Min, HUANG Da-fang |
1、Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
2、State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of
Agricultural Sciences, Beijing 100193, P.R.China |
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摘要 Using linker peptide LP4/2A for multiple gene transformation is considered to be an effective method to stack or pyramid several traits in plants. Bacillus thuringiensis (Bt) cry gene and epsps (5-enolpyruvylshikimate-3-phosphate synthase) gene are two important genes for culturing pest-resistant and glyphosate-tolerant crops. We used linker peptide LP4/2A to connect the Bt cry1Ah gene with the 2mG2-epsps gene and combined the wide-used manA gene as a selective marker to construct one coordinated expression vector called p2EPUHLAGN. The expression vector was transferred into maize by Agrobacterium tumefaciens-mediated transformation, and 60 plants were obtained, 40% of which were positive transformants. Molecular detection demonstrated that the two genes in the fusion vector were expressed simultaneously and spliced correctly in translation processing; meanwhile bioassay detection proved the transgenic maize had preferable pest resistance and glyphosate tolerance. Therefore, linker peptide LP4/2A provided a simple and reliable strategy for producing gene stacking in maize and the result showed that the fusion gene transformation system of LP4/2A was feasible in monocot plants.
Abstract Using linker peptide LP4/2A for multiple gene transformation is considered to be an effective method to stack or pyramid several traits in plants. Bacillus thuringiensis (Bt) cry gene and epsps (5-enolpyruvylshikimate-3-phosphate synthase) gene are two important genes for culturing pest-resistant and glyphosate-tolerant crops. We used linker peptide LP4/2A to connect the Bt cry1Ah gene with the 2mG2-epsps gene and combined the wide-used manA gene as a selective marker to construct one coordinated expression vector called p2EPUHLAGN. The expression vector was transferred into maize by Agrobacterium tumefaciens-mediated transformation, and 60 plants were obtained, 40% of which were positive transformants. Molecular detection demonstrated that the two genes in the fusion vector were expressed simultaneously and spliced correctly in translation processing; meanwhile bioassay detection proved the transgenic maize had preferable pest resistance and glyphosate tolerance. Therefore, linker peptide LP4/2A provided a simple and reliable strategy for producing gene stacking in maize and the result showed that the fusion gene transformation system of LP4/2A was feasible in monocot plants.
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Received: 17 February 2014
Accepted:
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| Fund: The authors gratefully acknowledge the financial support of the National Natural Science Foundation of China (30771383), the Genetically Modified Organisms Breeding Major Projects, China (2013ZX08003-001) and the National Basic Research Program of China (2009CB118902). |
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Corresponding Authors:
LANG Zhi-hong, Tel/Fax: +86-10-82109857,E-mail: langzhihong@caas.cn; HUANG Da-fang, Tel/Fax: +86-10-82109857, E-mail: huangdafang@caas.cn
E-mail: langzhihong@caas.cn;huangdafang@caas.cn
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| About author: SUN He, Mobile: 15010110522, E-mail: ndsh@163.com; |
Cite this article:
SUN He, LANG Zhi-hong, LU Wei, ZHANG Jie, HE Kang-lai , ZHU Li, LIN Min, HUANG Da-fang.
2015.
Developing transgenic maize (Zea mays L.) with insect resistance and glyphosate tolerance by fusion gene transformation. Journal of Integrative Agriculture, 14(2): 305-313.
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