A method for genetic transformation of Penicillium italicum mediated by Agrobacterium tumefaciens
A genetic transformation method, the technology of Agrobacterium tumefaciens, applied in the direction of using vectors to introduce foreign genetic material, recombinant DNA technology, etc., can solve the problems of agricultural production economic loss, fruit rot, etc., and achieve high transformation rate, stable transformation, and simple operation Effect
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[0033] The present invention will be described in detail below by way of examples.
[0034] 1. Experimental materials
[0035] 1.1 Strains and vectors
[0036] Agrobacterium tumefaciens GV3101, Agrobacterium tumefaciens AGL-1, Escherichia coli DH5α
[0037] The pMD19-T vector, plasmid pSKH, and plasmid pCAMBIA3300 were used to construct the knockout vector pacC3300
[0038] 1.2 Medium
[0039] Potato Dextrose Agar Medium (PDA): Potato 200g, take the filtrate after boiling, add 10g of sucrose, 20g of agar, for the cultivation of Penicillium italicum.
[0040] Beef extract peptone liquid medium (LB): tryptone 10g, yeast extract powder 5g, NaCl 10g, adjust pH to 7.4 with NaOH, add ddH2O to 1L.
[0041] Beef peptone solid medium (LBA): add 20g agar on the basis of LB medium
[0042] Table 1 Medium composition table
[0043]
[0044] Note: Glucose, AS and MES need to be sterilized by filtration.
[0045] 2. Construction of knockout vector
[0046] 2.1 Construction of the...
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