Yamaguchi R, Matsuo K, Yamazaki A, Kagawa H, Nagai S, Yamada T
Central Research Laboratories of Ajinomoto Co. Inc., Kawasaki, Japan.
Can J Microbiol. 1991 Jan;37(1):7-13. doi: 10.1139/m91-002.
The gene coding for the 12-kDa protein (MPB57) of Mycobacterium bovis BCG has recently been cloned and sequenced (R. Yamaguchi, K. Matsuo, A. Yamazaki, S. Nagai, K. Terasaka, and T. Yamada. 1988. FEBS Lett. 240: 115-117). To map linear B-cell epitopes by beta-galactosidase fusion proteins, we have constructed convenient vectors (pUR278S, pUR288S, and pUR289S) with the SmaI site. Based on recognition by polyclonal antibodies, two epitope regions on the MPB57 protein were identified, both of which corresponded to the amino acid sequences Glu20 to Val45 (26 residues, epitope I region) and Ile78 to Leu86 (9 residues, epitope II). Complementary oligonucleotides encoding epitope II were synthesized, polymerized by a ligase reaction, inserted into the native MPB57 protein gene, and expressed in Escherichia coli, giving rise to epitope-inserted proteins. Their stability and potential uses are described.
编码卡介苗牛分枝杆菌12 kDa蛋白（MPB57）的基因最近已被克隆和测序（R. 山口、K. 松尾、A. 山崎、S. 永井、K. 寺坂和T. 山田。1988年。《欧洲生物化学学会联合会快报》240：115 - 117）。为了通过β-半乳糖苷酶融合蛋白绘制线性B细胞表位图谱，我们构建了带有SmaI位点的便捷载体（pUR278S、pUR288S和pUR289S）。基于多克隆抗体的识别，在MPB57蛋白上鉴定出两个表位区域，它们都对应于氨基酸序列Glu20至Val45（26个残基，表位I区域）和Ile78至Leu86（9个残基，表位II）。合成了编码表位II的互补寡核苷酸，通过连接酶反应进行聚合，插入到天然MPB57蛋白基因中，并在大肠杆菌中表达，产生了表位插入蛋白。描述了它们的稳定性和潜在用途。
