HIGHLY CONSERVED GLUCOSE-REGULATED PROTEIN IN HAMSTER AND CHICKEN-CELLS - PRELIMINARY CHARACTERIZATION OF ITS CDNA CLONE
HIGHLY CONSERVED GLUCOSE-REGULATED PROTEIN IN HAMSTER AND CHICKEN-CELLS - PRELIMINARY CHARACTERIZATION OF ITS CDNA CLONE
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DOI：
10.1073/pnas.78.8.4922
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
SCHARFF, D
中科院分区：
文献类型：
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作者：
LEE, AS;DELEGEANE, A;SCHARFF, D
来源于中国仓鼠成纤维细胞的温度敏感突变体K12在非允许温度（40.5 ℃）下温育时过量产生MW为94，000、78，000和58，000的3种特异性蛋白质。C）。这些蛋白质先前被鉴定为葡萄糖调节蛋白，类似于当细胞缺乏葡萄糖时在鸡胚成纤维细胞中观察到的那些蛋白质。从仓鼠K12细胞系和鸡胚成纤维细胞中分离的78，000 MW蛋白质在二维等电聚焦凝胶中具有相同的电泳迁移率和几乎相同的肽图谱。这些蛋白质与之前描述的动物细胞热休克蛋白不同。通过使用从在40.5 ℃温育的仓鼠K12细胞中提取的RNA构建c[互补]DNA克隆文库。C.优先与由RNA制备的cDNA在40.5 °杂交的克隆。C被选中。通过使用杂交选择技术，然后在体外翻译，一个cDNA克隆含有2550个核苷酸的插入编码仓鼠MW 78，000蛋白质被确定。
A temperature-sensitive mutant K12 derived from a Chinese hamster fibroblast overproduces 3 specific proteins of MW 94,000, 78,000 and 58,000 when incubated at the nonpermissive temperature (40.5.degree. C). These proteins were previously identified as glucose-regulated proteins similar to those observed in chicken embryo fibroblasts when the cells are starved of glucose. The 78,000 MW proteins isolated from the hamster K12 cell line and from chicken embryo fibroblasts have identical electrophoretic mobilities in 2-dimensional isoelectric focusing gels and nearly identical peptide maps. These proteins are different from heat-shock proteins previously described for animal cells. A library of c[complementary]DNA clones was constructed by using the RNA extracted from the hamster K12 cells incubated at 40.5.degree. C. Clones that hybridize preferentially with cDNA made from RNA at 40.5.degree. C were selected. By using the hybrid-selection technique, followed by in vitro translation, a cDNA clone containing a 2550-nucleotide insert coding for the hamster MW 78,000 protein was identified.