Agrobacterium-mediated spirodela polyrhiza stable transformation system establishment method
An Agrobacterium-mediated and stable technology, applied in the fields of molecular biology and genetic engineering, can solve problems such as unreported
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[0065] A method for the establishment of a stable transformation system of Zipinkia gene mediated by Agrobacterium {auxin reporter gene (DR5::GUS) and cytokinin reporter gene (TCS::GUS)}. DR5::GUS is a synthetic auxin-responsive construct (Ulmasov et al., 1997). DR5 is composed of multiple 11-base repeat sequences, including the auxin response element TGTCTC sequence, which is fused with the GUS reporter gene to mark the auxin response; TCS::GUS is also a synthetic construction that marks the distribution of cytokinins, and is produced by cells The mitogen B-type response regulatory site and the minimal 35S promoter (minimal promoter), which are fused with the GUS reporter gene (Müller and Sheen, 2008). The specific implementation steps are as follows:
[0066] 1. Transformation of plant expression vector pKGWFS7.0 in Escherichia coli (DH5α):
[0067] The plant expression vector pKGWFS7.0 (purchased from Shanghai Maiqi Biotechnology Co., Ltd., the same below) with auxin repo...
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