Plasmid vector pET-mChBac75Na, strain of the bacterium Eschrichia coli BL (DE3 / pET-mChBac75Na for expression of the antimicrobial peptide of minibactenecin ChBac7Na and a method for producing said peptide
Федеральное государственное бюджетное учреждение науки Институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук (ИБХ РАН) (RU)
发明人:
Баландин Сергей Владимирович (RU),Болосов Илья Александрович (RU),Пантелеев Павел Валерьевич (RU),Кокряков Владимир Николаевич (RU),Шамова Ольга Валерьевна (RU),Овчин
申请号：
RU2016123389
公开号：
RU2016123389A
申请日:
2016.06.14
申请国别(地区):
RU
年份:
2017
代理人:
摘要：
FIELD: biotechnology.SUBSTANCE: invention relates to recombinant production of ChBac7.5Nα minibactenecin antimicrobial peptide from Capra hircus goat leukocytes, which can find application in medical and veterinary practice as a broad-spectrum antibiotic. A pET-mChBac75Na plasmid vector is constructed for ChBac7.5Nα minibactenecin expression in Escherichia coli cells as part of His8-TrxL-mChBac75Nα hybrid protein. This vector is used to transform the parent Escherichia coli BL21 (DE3) strain, obtaining a producing strain of His8-TrxL-mChBac75Nα fusion protein. Further cultivation of producing strain cells is performed, lysis is carried out together with affinity purification of His8-TrxL-mChBac75Nα fusion protein on the metal chelate carrier under denaturing conditions, cleavage of His8-TrxL-mChBac75Nα fusion protein by cyanogen bromide in 6M guanidine hydrochloride, and final target peptide purification by the HPLC reverse phase method.EFFECT: invention provides biologically active minibactenecin pruduction using a simplified technology.3 cl, 3 dwg, 1 tbl, 5 ex
