Antimicrobial peptide as well as preparation method and application thereof
An antibacterial peptide and signal peptide technology, applied in the field of anti-nematode peptide and its preparation, can solve the problems of limited resources, easy loss of resistance, easy to be affected by the environment, etc.
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[0043] The method for preparing the antimicrobial peptide developed by the present invention comprises the following steps, (1) adopting the trizol method to extract the total RNA of the pepper leaf tissue, and adopting reverse transcriptase to obtain the first strand cDNA for subsequent use; the reverse transcription used Enzyme: First-Strand Synthesis Kit (Invitrogen).
[0044] (2) According to the sequence of the known antimicrobial peptide Snakin gene, design upstream primers and downstream primers, use the first-strand cDNA as a template, clone the gene coding sequence without signal peptide in the antimicrobial peptide Snakin through PCR technology, and perform sequencing to obtain the complete The long gene sequence is recorded as the Sn gene sequence; the upstream primer sequence is 5'ATT CAA ACT GAT CAT GTT GC3', and the downstream primer sequence is 5'TTA AGG GCA TTT GCG TCT3';
[0045] (3) Cloning the Sn gene sequence obtained in step (2) into the prokaryotic expres...
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