A kind of disinfection preparation and disinfecting wet towel its method of preparation and application
A technology for disinfecting preparations and wet wipes, which is applied to the field of disinfecting preparations and disinfecting wet wipes containing antimicrobial peptides, and can solve the problems that cannot be completely overcome and the effective dose is low.
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Embodiment 1
[0039] Preparation and separation and purification of antimicrobial peptides
[0040] GK-1, GK-2 and GK-3 were prepared according to the above sequence, while cecropin A1 and buforin II were prepared as controls.
[0041] Sequence of cecropin A1 (see Morishima, I., etc., Comp. Biochem. Physiol., 1990, B 95(3), 551-554):
[0042] Arg Trp Lys Leu Phe Lys Lys Ile Glu Lys Val Gly Arg Asn Val Arg Asp Gly
[0043] Leu Ile Lys Ala Gly Pro Ala Ile Ala Val Ile Gly Gln Ala Lys Ser Leu
[0044] Sequence of buforin II (see Park, C.B., Biochem. Biophys. Res. Commun. 1996, 218(1), 408-413)
[0045] Thr Arg Ser Ser Arg Ala Gly Leu Gln Phe Pro Val Gly Arg Val His Arg Leu
[0046] Leu Arg Lys
[0047] In this example, solid-phase chemical synthesis was adopted, and the instrument used was the Pioneer polypeptide synthesizer produced by Applied Biosystems, Inc. of the United States. The synthesized peptide was sheared by high-concentration TFA, then purified by reverse column, and the puri...
Embodiment 2
[0064] Expression of antimicrobial peptide GK-1 gene in Escherichia coli
[0065] First design and synthesize the GK-1 gene encoding the antimicrobial peptide, clone it into the pGEX-4T1 vector (purchased from AmershamPharmcia Biotech), then transform Escherichia coli JM109, induce the expression of GST-GK-1 fusion protein by IPTG, and then undergo coagulation After enzymatic cleavage, the antimicrobial peptide GK-1 is obtained.
[0066] ATP, IPTG, T 4 polynucleotide kinase, T 4 DNA ligase, Klenow enzyme, and restriction endonuclease are all products of BIOLAB unless otherwise specified, the rubber tapping recovery kit is a product of Shanghai Sangon Company, and oligonucleotides are synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. Thrombin cleavage kit was purchased from SIGMA.
[0067] Molecular cloning operations such as DNA isolation, purification, PCR amplification, enzymatic hydrolysis, plasmid transformation of host bacteria, fragment recove...
Embodiment 3
[0077] Expression of Antimicrobial Peptide GK-1 Gene in Yeast Cells
[0078] ATP, IPTG, T 4 polynucleotide kinase, T 4 DNA ligase, Klenow enzyme, and restriction endonuclease are all products of BIOLAB unless otherwise specified, the rubber tapping recovery kit is a product of Shanghai Sangon Company, and oligonucleotides are synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. Thrombin cleavage kit was purchased from SIGMA.
[0079] The GK-1 gene encoding the antimicrobial peptide was designed and synthesized, which was digested with BamHI and ligated with the DNA sequence encoding GST, and then ligated into the plasmid pBluescriptSKII (purchased from Stratagene, USA) to transform Escherichia coli DH5α (purchased from Wuhan University strain Deposit Center), after the sequence of the extracted plasmid was confirmed to be correct by sequencing, the plasmid was digested with EcoRI and XhoI and connected to the pPIC9 vector (purchased from Invitrogen, USA...
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