Genetic transformation method of Eichhornia crassipes
A technology of water hyacinth and exogenous genes, which is applied in the field of genetic transformation of water hyacinth, can solve the problems of in-depth development and utilization limitations, difficult transgenic research, etc., and achieve the effects of easy mastery, popularization and application, simple operation and simple process
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Embodiment 1
[0050] Embodiment 1, transfer VP1 gene into water hyacinth
[0051] 1. Construction of plasmid pGII 0229-VP1
[0052] The construction process of plasmid pGII 0229-VP1 includes: artificially synthesizing the VP1 gene shown in Sequence 1, and making its two ends with the restriction site sequences of BamHI and SacI; digesting the synthetic gene and vector pBI121 with BamHI and SacI, and inserting The digested products were ligated to obtain the recombinant vector PBI121-VP1, in which the VP1 gene forms an expression cassette with the promoter 35S and the terminator nos. PBI121-VP1 and pGII 0229 were simultaneously digested with HindIII and EcoRI, ligated to obtain recombinant plasmids, transformed into Escherichia coli, resistance screening, liquid culture, plasmid extraction, enzyme digestion and sequencing identification. The result showed that the enzyme digestion result was correct; the measured VP1 gene sequence was shown in sequence 1, indicating that the constructed rec...
Embodiment 2
[0145] Implementation Example 2. Plant expression plasmid pGII 0229-GFP transforms water hyacinth.
[0146] 1. Construction of plasmid pGII 0229-GFP
[0147]The construction method of plasmid pGII 0229-GFP: the GFP gene shown in Sequence 2 is artificially synthesized, and its two ends are provided with XbaI and SacI restriction site sequences; the synthesized gene and vector pBI121 are digested with XbaI and SacI, and the enzyme The cut products were ligated to obtain the recombinant vector PBI121-GFP, in which the GFP gene forms an expression cassette with the promoter 35S and the terminator nos. Simultaneous digestion of pBI121-GFP and pGII 0229 with HindIII and EcoRI, ligation to obtain recombinant vector, transformation of Escherichia coli, resistance screening, liquid culture, extraction of plasmid, enzyme digestion and sequencing identification, the result of enzyme digestion was correct, and the measured GFP The gene sequence is shown in sequence 2, indicating that the...
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