A kind of genetic transformation method of Haematococcus pluvialis
A genetic transformation method, the technology of Haematococcus pluvialis, applied in the field of genetic transformation of Haematococcus pluvialls, can solve the problems of low positive transformation rate, low success rate, cumbersome operation, etc., achieve broad market prospects, long cycle, Avoiding the Effects of Cellular Stress Responses
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[0061] The present embodiment provides a genetic transformation method of Haematococcus pluvialis, comprising the following steps:
[0062] 1 Linearization of plasmids for transformation
[0063] The plasmid used in this example was pH124. The pH124 plasmid was extracted from Escherichia coli DH5α using Plasmid Mini Kit II (Omega Bio-tek, USA), the plasmid was linearized with restriction endonuclease Not I, and digested with Not I. The system is 30 μL, and specifically includes: 20 μL DNA solution, 2 μL Not I Fast Dicer (New England (NEB), USA), 3 μL Cutsmart Buffer (New England (NEB), USA), 5 μL water. The digestion temperature was 37° C. and the digestion time was 2 h to obtain a linearized pH124 plasmid with the DNA sequence shown in SEQ ID NO: 1, and the linearized plasmid concentration was about 1.0 μg / μL.
[0064] 2 Genetic transformation of Haematococcus pluvialis
[0065] (1) Collect Haematococcus pluvialis cells
[0066] The Haematococcus pluvialis cell selected i...
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