Zhou Liangfan, Lin Qingping, Li Baocun, Li Nannan, Zhang Shuangquan
Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing, 210046, China.
Biotechnol Lett. 2009 Mar;31(3):437-41. doi: 10.1007/s10529-008-9893-0. Epub 2008 Nov 27.
The antimicrobial peptide CM4 is a 35-residue cationic peptide. To explore a new approach for the expression and purification of CM4 in Escherichia coli, the CM4 gene was cloned into the vector pET32a to construct an expression vector pET32a-CM4. The fusion protein Trx-CM4, purified by Ni(2+)-chelating chromatography, was cleaved by hydroxylamine hydrochloride to release recombinant CM4. Purification of recombinant CM4 was achieved by reverse HPLC chromatography, and about 1.4 mg/l active recombinant CM4 with the purity more than 98% was obtained. The recombinant CM4 showed antimicrobial activities that were similar to synthetic one.
抗菌肽CM4是一种由35个氨基酸残基组成的阳离子肽。为探索在大肠杆菌中表达和纯化CM4的新方法，将CM4基因克隆到载体pET32a中构建表达载体pET32a-CM4。通过镍离子螯合层析纯化的融合蛋白Trx-CM4，用盐酸羟胺裂解以释放重组CM4。重组CM4通过反向高效液相色谱法进行纯化，获得了纯度大于98%、活性约为1.4mg/L的重组CM4。重组CM4表现出与合成CM4相似的抗菌活性。
