Fusion expression of cecropin B-like antibacterial peptide in Escherichia coli and preparation of its antiserum.
Antibacterial peptides have a broad range of antibacterial properties that makes them highly toxic for expression in Escherichia coli. For prepare an antiserum to detect these peptides, we developed a cecropin B mutant with a green fluorescent protein fusion partner resulting in high expression of a 37 kDa fusion peptide in E. coli with a yield of 7.9 mg/l culture medium after purification on Ni-IDA resin. Guinea pigs when immunized with the fusion peptide produced a specific antiserum which titers in excess of 1:25,600.
Yu
Fuxian
F
Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, Beijing, 100193, People's Republic of China.
Wang
Jigui
J
Zhang
Peng
P
Hong
Yan
Y
Liu
Weiquan
W
D000818:Animals; D000906:Antibodies / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D006168:Guinea Pigs; D019476:Insect Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D000906:Antibodies; D019476:Insect Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C031162:cecropin B protein, Insecta