Ishikawa K, Kurata N, Aoki H, Kato T, Marunouchi T
Division of Cell Biology, Fujita Health University, Aichi, Japan.
Hybridoma. 1991 Jun;10(3):357-68. doi: 10.1089/hyb.1991.10.357.
Valuable information about proto-oncogenes and their physiological functions has been obtained by studying their expression in normal cells. However, the protein product of the c-mos gene, the cellular homologue of the transforming gene (v-mos) of Moloney murine sarcoma virus, has not been detected in normal mouse cells or tissues. Here, we have constructed a v-mos expression vector, pRI-delta mos, which directs the synthesis of a truncated v-mos gene product, a protein A fusion protein. Using the truncated v-mos oncoprotein produced in Escherichia coli as immunogen, we prepared anti-v-mos monoclonal antibodies (MAbs). In immunoblotting assays, the MAb was reactive with v-mos oncoprotein and detected bands at 43 KDa or 39 kDa in the tissue extract of mouse testes or ovaries, respectively, in which the c-mos protooncogene mRNA is expressed. These results demonstrate that the v-mos MAb obtained is suitable for elucidating the physiological functions of v-mos gene product and may also be utilized to detect c-mos gene product at the cellular level.
通过研究原癌基因在正常细胞中的表达,已获得了有关原癌基因及其生理功能的宝贵信息。然而,莫洛尼鼠肉瘤病毒转化基因(v-mos)的细胞同源物c-mos基因的蛋白质产物,在正常小鼠细胞或组织中尚未检测到。在此,我们构建了一个v-mos表达载体pRI-δmos,它指导合成一种截短的v-mos基因产物,即一种蛋白A融合蛋白。以在大肠杆菌中产生的截短v-mos癌蛋白作为免疫原,我们制备了抗v-mos单克隆抗体(MAb)。在免疫印迹分析中,该单克隆抗体与v-mos癌蛋白发生反应,并且在小鼠睾丸或卵巢的组织提取物中分别检测到43 kDa或39 kDa的条带,其中c-mos原癌基因mRNA在这些组织中表达。这些结果表明,所获得的v-mos单克隆抗体适用于阐明v-mos基因产物的生理功能,并且也可用于在细胞水平检测c-mos基因产物。