Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, Beijing, 100193, People's Republic of China.
Biotechnol Lett. 2010 May;32(5):669-73. doi: 10.1007/s10529-009-0196-x. Epub 2010 Jan 3.
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.
抗菌肽具有广谱的抗菌特性,使其在大肠杆菌中表达时具有高度毒性。为了制备抗体制检测这些肽,我们开发了一种带有绿色荧光蛋白融合伴侣的抗菌肽突变体,在大肠杆菌中高表达了一个 37 kDa 的融合肽,经 Ni-IDA 树脂纯化后,产量为 7.9mg/L 培养基。用融合肽免疫豚鼠产生了效价超过 1:25600 的特异性抗血清。