A method for genetic transformation of Laminaria gametophytes mediated by Agrobacterium
A genetic transformation method, Agrobacterium-mediated technology, applied in biochemical equipment and methods, botany equipment and methods, fermentation, etc., can solve the problems of cumbersome detection process and low transformation efficiency, so as to improve the efficiency of genetic transformation and shorten the The effect of the conversion cycle
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[0043] (1) Preparation of infested materials: dry the species of kelp (the species of kelp collected from August to September 2014 with the number 201409006) and disperse it in the shade. After 4 hours, use the conditions of 4000rpm and 10min to enrich the zoospores. The culture medium dilutes the kelp zoospores so that the density of the kelp zoospores is 2×10 4 individual / mL to obtain the kelp zoospore suspension;
[0044] (2) Activation of Agrobacterium: use co-culture solution to cultivate and activate Agrobacterium halotolerance to OD=0.5; obtain culture solution of Agrobacterium halotolerance;
[0045] (3) Infection co-cultivation: centrifuge the salt-tolerant Agrobacterium culture solution activated in step (2) to remove the supernatant, and the precipitated bacteria are re-selected with the co-culture solution, and then added to the kelp zoospore suspension obtained in step (1). halotolerant Agrobacterium halotolerant to the initial OD value of 0.114-0.263, and use th...
Embodiment 1
[0049] Example 1: The Agrobacterium used for infection was Agrobacterium EHA105 transformed with the pBI221-BAR-GFP plasmid, and the initial OD value of the infection co-culture was 0.132. The microscopic observation of the infected co-culture and the observation results under the blue light of the fluorescence microscope are as follows: figure 1 shown.
Embodiment 2
[0050] Example 2: The Agrobacterium used for infection is Agrobacterium GV3101 transformed into pBI221-BAR-GFP, and the initial OD value of infection co-culture is 0.221. The microscopic observation of the infected co-culture and the observation results under the blue light of the fluorescence microscope are as follows: figure 1 shown.
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