Construction and expression of a novel bioactive IFN-alpha2b/CM4 fusion protein in Escherichia coli.
Human interferon alpha2b (IFN-alpha2b) is a pleiotropic cytokine used for the treatment of various cancers. Antibacterial peptide CM4 is a small peptide that can strongly inhibit many kinds of bacteria, fungi, and tumor cells, but it does no harm to normal cells. Here, we describe a protein expression system for the production of IFN-alpha2b/CM4 fusion protein in insoluble form in Escherichia coli, coupled to an efficient dialysis refolding and histidine-tag purification protocol. The expressed IFN-alpha2b/CM4 fusion protein not only displays significantly improved antitumor activity, but also retains the antibacterial-antifungal activity of CM4.
Li
Nan-Nan
NN
Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210046, China.
Liu
Ping
P
Chen
Su-Juan
SJ
Lin
Qing-Ping
QP
Zhou
Liang-Fan
LF
Zhang
Shuang-Quan
SQ
D000818:Animals; D000890:Anti-Infective Agents / Q000302:isolation & purification / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects; D045744:Cell Line, Tumor; D004847:Epithelial Cells / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics; D005658:Fungi / Q000187:drug effects; D006801:Humans; D002479:Inclusion Bodies; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D011994:Recombinant Proteins
D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D000970:Antineoplastic Agents; C534224:CM4 peptide, Bombyx mori; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins