Hartl M, Bister K
Institute of Biochemistry, University of Innsbruck, Austria.
Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11731-5. doi: 10.1073/pnas.92.25.11731.
We have analyzed differential gene expression in normal versus jun-transformed avian fibroblasts by using subtracted nucleic acid probes and differential nucleic acid hybridization techniques for the isolation of cDNA clones. One clone corresponded to a gene that was strongly expressed in a previously established quail (Coturnix japonica) embryo fibroblast line (VCD) transformed by a chimeric jun oncogene but whose expression was undetectable in normal quail embryo fibroblasts. Furthermore, the gene was expressed in quail or chicken fibroblast cultures that were freshly transformed by retroviral constructs carrying various viral or cellular jun alleles and in chicken fibroblasts transformed by the avian retrovirus ASV17 carrying the original viral v-jun allele. However, its expression was undetectable in a variety of established avian cell lines or freshly prepared avian fibroblast cultures transformed by other oncogenes or a chemical carcinogen. The nucleotide and deduced amino acid sequences of the cDNA clone were not identical to any sequence entries in the data bases but revealed significant similarities to avian beta-keratin genes; the highest degree of amino acid sequence identity was 63%. The gene, which we termed bkj, may represent a direct or indirect target for jun function.
我们通过使用扣除核酸探针和差异核酸杂交技术来分离cDNA克隆，分析了正常与jun转化的禽成纤维细胞中的差异基因表达。一个克隆对应一个基因，该基因在先前建立的由嵌合jun癌基因转化的鹌鹑（日本鹌鹑）胚胎成纤维细胞系（VCD）中强烈表达，但在正常鹌鹑胚胎成纤维细胞中未检测到其表达。此外，该基因在由携带各种病毒或细胞jun等位基因的逆转录病毒构建体新转化的鹌鹑或鸡成纤维细胞培养物中以及由携带原始病毒v-jun等位基因的禽逆转录病毒ASV17转化的鸡成纤维细胞中表达。然而，在各种已建立的禽细胞系或由其他癌基因或化学致癌物新制备的禽成纤维细胞培养物中未检测到其表达。该cDNA克隆的核苷酸和推导的氨基酸序列与数据库中的任何序列条目都不相同，但与禽β-角蛋白基因显示出显著的相似性；氨基酸序列同一性的最高程度为63%。我们将该基因命名为bkj，它可能代表jun功能的直接或间接靶点。
