CRISPR/Cpf1 gene editing method applicable to marine crustacean mitochondrion genome
A mitochondrial genome and gene editing technology, applied in the field of molecular biology, can solve unseen problems and achieve the effect of simple operation and good editing effect
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[0017] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art.
[0018] The implementation of the present invention will be further described in detail below in conjunction with specific examples of application in Portunus trituberculatus. It should be understood that the examples are only for illustrating the present invention and should not and will not be used to limit the scope of the present invention. Various procedures and methods not described in detail in the following examples are conventional methods well known in the art.
[0019] 1. Construction of plasmid vector
[0020] Since there is no suitable restriction endonuclease available in the NLS sequence region of pY094 plasmid Cpf1, it is necessary to introduce two enzyme cutting site regions, XcmI (position: 4870bp) and BamHI (position 5201), specifically as follows, manually 合成“5'CACCATGGTGGCCCTGATCCGCAGCGTGCTGCAGATGCGGAACT...
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