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在大肠杆菌中融合表达抗菌肽 Cecropin B 样并制备其抗血清。

Fusion expression of cecropin B-like antibacterial peptide in Escherichia coli and preparation of its antiserum.

机构信息

Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, Beijing, 100193, People's Republic of China.

出版信息

Biotechnol Lett. 2010 May;32(5):669-73. doi: 10.1007/s10529-009-0196-x. Epub 2010 Jan 3.

Abstract

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.

摘要

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。

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