Yang H W, Hemmi H, Ikeda H, Kato K, Tsuchida Y
Department of Pediatric Surgery, Faculty of Medicine, University of Tokyo, Bunkyo-ku, Tokyo 113-8655, Japan.
Oncol Rep. 1999 Jan-Feb;6(1):107-11.
The importance of determining the N-Myc protein has been emphasized in neuroblastoma. We attempted to obtain a water-soluble N-Myc protein, and an antibody highly specific for the N-Myc protein. A plasmid was constructed from partial exons 2 and 3 of the N-myc gene, and expressed in Escherichia coli by isopropyl-beta,-D-thiogalactopyranoside. Newly obtained N-Myc (rN-Myc) was water-soluble with a M.W. of 38 kDa. An N-Myc-specific peptide, GVAPPRPGG RQTSGGDH, was used to raise an antibody. Specificity of the obtained antibody was confirmed and we found the rN-Myc protein reacts positively with the anti-N-Myc IgG raised here. The rN-Myc protein and the anti-N-Myc IgG obtained are expected to be used in an ELISA for N-Myc.
确定N-Myc蛋白在神经母细胞瘤中的重要性已得到强调。我们试图获得一种水溶性N-Myc蛋白以及对N-Myc蛋白具有高度特异性的抗体。从N-myc基因的部分外显子2和3构建了一个质粒，并通过异丙基-β-D-硫代半乳糖苷在大肠杆菌中表达。新获得的N-Myc（rN-Myc）是水溶性的，分子量为38 kDa。使用一种N-Myc特异性肽GVAPPRPGG RQTSGGDH来制备抗体。所获得抗体的特异性得到了证实，并且我们发现rN-Myc蛋白与在此制备的抗N-Myc IgG呈阳性反应。预期所获得的rN-Myc蛋白和抗N-Myc IgG可用于N-Myc的酶联免疫吸附测定（ELISA）。
