Artificially synthesized herbicide-resistant protein
A protein and protein technology, applied in the biological field, can solve the problems of cotton that have not been reported yet
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Embodiment 1
[0039] Embodiment 1, EPSPS fusion gene TS-EPSPS
[0040] All the following gene manipulation methods were carried out with reference to the description in "Molecular Cloning" (Sambrook J et al., 1989); the restriction endonucleases and other tool enzymes used were all purchased from NEB Company.
[0041] 1. Acquisition of EPSPS fusion gene TS-EPSPS and fusion gene ACP1-EPSPS
[0042] 1. EPSPS codon optimization transformation and chemical synthesis
[0043] The codon optimization strategies for the herbicide-resistant protein coding gene (EPSPS gene) sequence of wild-type Agrobacterium agrobacterium CP4 mainly include: the application of plant-favored codons; the transformation of unstable sequences; the increase of GC content, etc. The G+C content of the wild-type CP4 strain EPSPS gene is very low, and the A+T content is very high. On the one hand, if these sequences are introduced into the plant genome, they may be mistaken for plant gene control sequences (A+ of known sequ...
Embodiment 2
[0064] Embodiment 2, the acquisition and performance analysis of transgenic cotton
[0065] 1. Obtaining genetically modified cotton
[0066] 1. Obtaining genetically modified cotton
[0067] The recombinant bacteria LBA4404 / pBI121-EPSPS, LBA4404 / pBI121-ACP1-EPSPS and LBA4404 / pBI121-TS-EPSPS transformed into cotton (No. Zhongmian No. 35, hereinafter also referred to as wild-type cotton) in Example 1, specifically transformed Method and screening reference of transgenic cotton plants (Wu JH, Zhang XL, Nie YC, Luo, XY. High-efficiency.transformation of Gossypium hirsutum embryogenic calli.mediated by Agrobacterium tumefaciens and regeneration of insect-resistant plants.2005, Plant Breed.124 : 142-146), to obtain T0 generation EPSPS cotton, T0 generation transfer TS-EPSPS cotton and T0 generation transfer ACP1-EPSPS cotton.
[0068] The same method was used to transform the empty vector pBI121 into wild-type cotton to obtain the T0 generation transformed cotton (EV).
[0069] ...
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