Alvarez A M, Fukuhara E, Nakase M, Adachi T, Aoki N, Nakamura R, Matsuda T
Department of Applied Biological Sciences, School of Agricultural Sciences, Nagoya University, Japan.
Biosci Biotechnol Biochem. 1995 Jul;59(7):1304-8. doi: 10.1271/bbb.59.1304.
Four rice seed proteins encoded by cDNAs belonging to the alpha-amylase/trypsin inhibitor gene family were overexpressed as TrpE-fusion proteins in E. coli. The expressed rice proteins were detected by SDS-PAGE as major proteins in bacterial cell lysates. Western blot analyses showed that all the recombinant proteins were immunologically reactive to rabbit polyclonal antibodies and to a mouse monoclonal antibody (25B9) specific for a previously isolated rice allergen of 16 kDa. Some truncated proteins from deletion mutants of the cDNAs retained their reactivity to the specific antibodies. These results suggest that the cDNAs encode potential rice allergens and that some epitopes of the recombinant proteins are still immunoreactive when they are expressed as their fragments.
由属于α-淀粉酶/胰蛋白酶抑制剂基因家族的cDNA编码的四种水稻种子蛋白在大肠杆菌中作为TrpE融合蛋白过表达。通过SDS-PAGE检测到表达的水稻蛋白是细菌细胞裂解物中的主要蛋白。蛋白质印迹分析表明，所有重组蛋白对兔多克隆抗体和对先前分离的16 kDa水稻过敏原具有特异性的小鼠单克隆抗体（25B9）具有免疫反应性。来自cDNA缺失突变体的一些截短蛋白保留了它们对特异性抗体的反应性。这些结果表明，这些cDNA编码潜在的水稻过敏原，并且当重组蛋白以片段形式表达时，它们的一些表位仍然具有免疫反应性。
