A broad-spectrum and high-efficiency antimicrobial peptide pb-cath-oh1 and its gene, preparation method and application
An antimicrobial peptide, coding gene technology, applied in the field of biomedicine, to achieve the effects of low cytotoxicity, strong antibacterial activity, convenient chemical synthesis and genetic engineering preparation
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[0020] Specific embodiments of the present invention will be described in detail below in conjunction with technical solutions, but the content of the present invention is not limited thereto.
[0021] Strictly follow the kit instructions, using RNeasy AxyPrep TM Multisource Total RNA Miniprep Kit (Qiagen, union city, CA, USA) was used to extract the total RNA from Burmese python lung tissue, and then use Creator TM SMART TMThe cDNA Library Construction Kit was used to construct the cDNA library of Burmese python lung tissue. Use the PowerScript Reverse Transcriptase in the kit to reverse transcribe and synthesize the first strand of cDNA. The primers are:
[0022] Forward SMARTⅣOligonucleotide primer:
[0023] 5'–AAGCAGTGGTATCAACGCAGAGTGGCCATTACGGCCGGG–3'
[0024] Reverse CDSⅢ / 3'PCR primer: 5'–ATTCTAGAGGCCGAGGCGGCCGACATG-d(T) 30 N -1 N–3’ (N=A, C, G, or T; N -1 = A, G, or C).
[0025] Use Advantage DNA Polymerase to synthesize the second strand of cDNA. The primer i...
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