Introduction and expression of a fetal human globin gene in mouse fibroblasts.

N Hsiung; R S Roginski; P Henthorn; O Smithies; R Kucherlapati; A I Skoultchi

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Introduction and expression of a fetal human globin gene in mouse fibroblasts.

机译：胎儿人球蛋白基因在小鼠成纤维细胞中的导入和表达。

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An 8.5-kilobase segment of cloned human DNA including the complete G gamma-globin gene was introduced into LMTK- cells by the calcium phosphate precipitation method in the presence or absence of carrier DNA. Transfectants containing one or more copies of intact G gamma-globin genes were obtained either by ligation of the human DNA segment to a plasmid containing the herpes simplex virus thymidine kinase gene or by nonligated cotransfer. The integrity of the integrated gamma-globin gene was established by Southern blotting experiments. Expression of the herpes simplex virus thymidine kinase and human gamma-globin genes was evaluated by Northern blotting and solution hybridization. Of 23 transfectants analyzed, 21 produced a 9S gamma-globin RNA migrating like authentic gamma-globin mRNA on denaturing agarose gels. The gamma-globin RNA is polyadenylated and present in the cytoplasm of the transfected cells; it accumulates to a level 10 times that of thymidine kinase mRNA, or about 5 to 50 molecules per transfected cell. By using plasmids in which the gamma-gene is inserted in either transcriptional orientation with respect to the thymidine kinase gene, it was possible to show that transcription occurred from the gamma-gene promoter.

机译：在存在或不存在载体DNA的情况下，通过磷酸钙沉淀法将包括完整的Gγ-球蛋白基因的克隆的人类DNA的8.5千碱基片段引入LMTK细胞。通过将人DNA片段连接到含有单纯疱疹病毒胸苷激酶基因的质粒上或通过未连接的共转移来获得含有一个或多个完整Gγ-珠蛋白基因拷贝的转染子。通过Southern印迹实验确定了整合的γ-珠蛋白基因的完整性。通过Northern印迹和溶液杂交评估单纯疱疹病毒胸苷激酶和人γ-珠蛋白基因的表达。在分析的23种转染子中，有21种产生了9Sγ-珠蛋白RNA，就像变性的γ-珠蛋白mRNA在变性琼脂糖凝胶上一样迁移。 γ-珠蛋白RNA是聚腺苷酸化的，存在于转染细胞的细胞质中。它的积累水平是胸苷激酶mRNA的10倍，或每个转染细胞约5至50个分子。通过使用其中γ基因相对于胸苷激酶基因以任一转录方向插入的质粒，有可能表明转录是从γ基因启动子发生的。

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《Molecular and Cellular Biology》 |1982年第4期|共11页
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N Hsiung; R S Roginski; P Henthorn; O Smithies; R Kucherlapati; A I Skoultchi;
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中图分类细胞生物学;
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1. BP1 motif in the human beta-globin promoter affects beta-globin expression during embryonic/fetal erythropoiesis in transgenic mice bearing the human beta-globin gene. [J] . Zoueva OP, Garrett LJ, Bodine D, Blood cells, molecules and diseases . 2008,第3期

机译：人β-珠蛋白启动子中的BP1基序会在携带人β-珠蛋白基因的转基因小鼠的胚胎/胎儿红细胞生成过程中影响β-珠蛋白的表达。
2. Developmental regulation of fetal to adult globin gene switching in human fetal erythroid × mouse erythroleukemia cell hybrids☆ [J] . Tariq Enver, Martha Brice, Joyce Karlinsey, Developmental biology . 1991,第1期

机译：人胎儿红系×小鼠红白血病细胞杂交物中胎儿向成年球蛋白基因转换的发育调控☆
3. Persistent fetal gamma-globin expression in adult transgenic mice following deletion of two silencer elements located 3' to the human Agamma-globin gene. [J] . Molecular medicine. . 2009,第11a12期

机译：在缺失位于人类Agamma-globin基因3'的两个沉默子元件后，成年转基因小鼠中胎儿的γ-globin持续表达。
4. Analysis of IDO Gene Expressions in Mouse Fetal Liver Mesenchymal Stem Cells after Cryopreservation Using Different Programs [C] . Dimitrov A., Bondarovich N., Chelombitko O., 13th Cryogenics 2014 . 2014

机译：冷冻保存不同程序后小鼠胎儿肝间充质干细胞中IDO基因表达的分析
5. GENE CONVERSION IN THE EVOLUTION OF THE HUMAN FETAL GAMMA GLOBIN GENES [D] . BLECHL, ANN ELIZABETH. 1981

机译：人类胎儿丙种球蛋白基因进化中的基因转化
6. Introduction and expression of a fetal human globin gene in mouse fibroblasts. [O] . N Hsiung, R S Roginski, P Henthorn, 1982

机译：胎儿人球蛋白基因在小鼠成纤维细胞中的导入和表达。
7. Introduction and expression of a fetal human globin gene in mouse fibroblasts. [O] . Hsiung, N, Roginski, R S, Henthorn, P, 1982

机译：胎儿人球蛋白基因在小鼠成纤维细胞中的导入和表达。

Introduction and expression of a fetal human globin gene in mouse fibroblasts.

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