Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche, G. Salvemini 1, 06126, Perugia, Italy.
Protein J. 2010 Apr;29(3):153-60. doi: 10.1007/s10930-010-9228-6.
Bovine leukaemia virus (BLV) causes disease in cattle and it is related to human T lymphotrofic viruses HTLV-1 and HTLV-2. The objective of this study was to express and purify deleted and stable forms of the gp51 envelope glycoprotein of BLV using a baculovirus system. Two forms of the gp51 were synthesised: one comprised the gp51 N-terminal 174 amino acids and a single 6xHis tag (Delta(175-268)gp51-His) and the second form contained the same amino acid sequence and a C-terminal Strep-tag II in addition to the 6xHis tag (Delta(175-268)gp51-STH). The two proteins were expressed and purified by immobilized metal-affinity chromatography (IMAC) or by Strep-Tactin column. The Strep-Tactin technology was more efficient than IMAC method and achieved a high pure recombinant deleted gp51. In addition the Delta(175-268)gp51-STH protein was further concentrated by IMAC. This purified antigen could be used for the isolation of immunoreactive molecules and to develop a competitive ELISA test.
牛白血病病毒（BLV）可引起牛的疾病，并且与人类 T 淋巴细胞白血病病毒 HTLV-1 和 HTLV-2 有关。本研究的目的是使用杆状病毒系统表达和纯化 BLV 的缺失和稳定形式的 gp51 包膜糖蛋白。合成了两种形式的 gp51：一种包含 gp51 N 端 174 个氨基酸和单个 6xHis 标签（Delta(175-268)gp51-His），第二种形式包含相同的氨基酸序列和 C 端 Strep-tag II 以及 6xHis 标签（Delta(175-268)gp51-STH）。这两种蛋白质通过固定化金属亲和层析（IMAC）或 Strep-Tactin 柱进行表达和纯化。Strep-Tactin 技术比 IMAC 方法更有效，可实现高纯度的重组缺失 gp51。此外，Delta(175-268)gp51-STH 蛋白通过 IMAC 进一步浓缩。这种纯化的抗原可用于分离免疫反应性分子，并开发竞争性 ELISA 测试。
