Cloning method of kosteletzkya virginica KvNAC gene and construction method of expression vector of kosteletzkya virginica KvNAC gene
A cloning method and gene technology, which are applied in the field of KvNAC gene cloning and expression vector construction of seaside mallow, can solve the problems of little research on the molecular mechanism of salt resistance and no KvNAC gene found.
Inactive Publication Date: 2020-11-10
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Problems solved by technology
The invention discloses a cloning method of a kosteletzkya virginica KvNAC gene and a construction method of an expression vector of the kosteletzkya virginica KvNAC gene. The cloning method comprisesthe steps of extracting total RNA of kosteletzkya virginica, performing cloning and sequence analyzing of the kosteletzkya virginica KvNAC gene, carrying out an RT-PCR reaction, and performing sequencing to obtain a full-length gene segment of the KvNAC gene; according to the obtained full length of the KvNAC gene, designing a primer for separation to obtain an ORF sequence of the gene; selectinghigh-fidelity Pfu DNA polymerase for PCR amplification to obtain a target segment; and taking 4 [mu]L of a PCR product and 1 [mu]L of a pEASY-Blunt Cloning Vector to carry out a ligation reaction soas to construct a recombinant cloning vector. The method is simple, convenient and effective; Isolation and cloning of the gene are realized in a new salt-resistant species, total RNA is successfullyextracted from the gene, and a spatial expression mode of the gene in a plant body is realized. The construction of the expression vector lays a foundation for the research of gene functions.
Microbiological testing/measurementFermentation +4
Cloning vectorSequence analysis +8
- Experimental program(1)
- Effect test(1)
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