Transient expression of genes introduced into cultured cells of Drosophila.
Transient expression of genes introduced into cultured cells of Drosophila.
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导入果蝇培养细胞的基因瞬时表达。
DOI：
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发表时间：
1983
影响因子：
11.1
通讯作者：
Igor B. Dawid
中科院分区：
文献类型：
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作者：
P. P. D. Nocera;Igor B. Dawid
将编码细菌氯霉素乙酰转移酶（CAT；乙酰辅酶A：氯霉素 3-O-乙酰转移酶，EC 2.3.1.28）的序列置于果蝇热休克蛋白 70（hsp 70）或 copia 启动子控制下的重组质粒已引入两种果蝇物种（黑腹果蝇的 Schneider II 系和D. immigrans）作为钙-磷酸盐复合物。转染后 1-2 天内，在暴露于任一 CAT 重组体的细胞中检测到有功能的 CAT 酶。细菌信息的表达取决于果蝇启动子的活性，因为果蝇DNA片段与CAT编码序列反向融合的质粒不支持CAT酶活性的合成。当细胞维持在室温下时，在用 hsp-cat 重组体转化的细胞中检测到低水平的 CAT 活性和杂合 mRNA，并且在短暂暴露于 37°C 后，mRNA 水平和 CAT 活性均大幅增加。hsp-cat mRNA 具有与真实果蝇 hsp 70 信使相同的 5' 末端。这些实验记录了在果蝇培养细胞中引入和表达分离基因的实用系统。
Recombinant plasmids in which the sequence encoding the bacterial chloramphenicol acetyltransferase (CAT; acetyl-CoA:chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28) has been placed under the control of Drosophila heat shock protein 70 (hsp 70) or copia promoters have been introduced into cultured cells of two Drosophila species (Schneider II line of Drosophila melanogaster and D. immigrans) as calcium-phosphate complexes. Within 1-2 days after transfection functional CAT enzyme was detected in cells exposed to either CAT recombinant. The expression of the bacterial information depends on the activity of the Drosophila promoters because plasmids in which the Drosophila DNA fragments were fused to the CAT coding sequence in inverted orientation did not support the synthesis of CAT enzyme activity. Low levels of CAT activity and of hybrid mRNA were detected in cells transformed with hsp-cat recombinants when the cells were maintained at room temperature, and both mRNA levels and CAT activity increased substantially after a brief exposure to 37 degrees C. hsp-cat mRNA has the same 5' terminus as authentic Drosophila hsp 70 messenger. These experiments document a practical system for the introduction and expression of isolated genes in cultured cells of Drosophila.