Engqvist Martin K M, Maurino Veronica G
Department of Biology and Biological Engineering, Division of Systems and Synthetic Biology, Chalmers University of Technology, Göteborg, Sweden.
Institute of Developmental and Molecular Biology of Plants, Plant Molecular Physiology and Biotechnology Group, Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich-Heine-Universität, Universitätsstraße 1, 40225, Düsseldorf, Germany.
Methods Mol Biol. 2017;1653:137-155. doi: 10.1007/978-1-4939-7225-8_10.
The introduction of two alternative glycolate catabolic pathways in the chloroplasts of Arabidopsis thaliana rendered plants with increased biomass. To introduce these synthetic pathways, the selected genes were stepwise integrated in the nuclear genome of wild-type plants. These plants were transformed by Agrobacterium tumefaciens carrying the binary vectors using the floral dip method. Selection of transformants was conducted using different selection agents and the expression of the transgenes was confirmed by PCR and enzyme activity measurements.
在拟南芥叶绿体中引入两条替代的乙醇酸分解代谢途径，使植株生物量增加。为引入这些合成途径，将选定的基因逐步整合到野生型植物的核基因组中。使用农杆菌介导的花浸染法，用携带双元载体的根癌农杆菌对这些植物进行转化。使用不同的选择剂对转化体进行筛选，并通过PCR和酶活性测量来确认转基因的表达。
