MICROINJECTION STUDIES OF DUCK GLOBIN MESSENGER-RNA TRANSLATION IN HUMAN AND AVIAN CELLS
MICROINJECTION STUDIES OF DUCK GLOBIN MESSENGER-RNA TRANSLATION IN HUMAN AND AVIAN CELLS
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DOI：
10.1016/0092-8674(76)90136-7
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发表时间：
1976-01-01
期刊：
影响因子：
64.5
通讯作者：
ALLFREY, VG
中科院分区：
文献类型：
--
作者：
STACEY, DW;ALLFREY, VG
显微注射技术用于研究活细胞中异特异性mRNA的翻译。将鸭珠蛋白mRNA以多聚体、信使核糖核蛋白（mRNP）颗粒或游离mRNA的形式注射到人源细胞（人宫颈癌细胞）中，采用特异性免疫荧光技术检测禽珠蛋白随后的形态和积累情况。注射后至少25小时，高比例的珠蛋白在HeLa细胞内积累，表明异特异性珠蛋白mRNA在细胞内的稳定性，以及最初与更复杂结构相关的mRNA可以在HeLa细胞质中翻译。在类似的实验中，鸭网状细胞多体在鸭胚胎细胞中被翻译，而在鸡胚胎肝细胞中没有被翻译。后者确实经受住了显微注射过程，因为它们对α合成的诱导剂和抑制剂有正常反应的能力。-氨基乙酰丙酸合成酶注射鸭Hb的HeLa细胞在注射后以正常或接近正常的速率分裂，但其珠蛋白含量减少，而接受鸭网状细胞多体、mRNP颗粒或游离mRNA的细胞的珠蛋白含量增加。
Microinjection techniques were used for the study of the translation of heterospecific mRNA in living cells. Duck globin mRNA, in the form of polysomes, messenger ribonucleoprotein [mRNP] particles or free mRNA, were injected into cells of human origin [human cervical carcinoma cells], and the subsequent appearance and accumulation of avian globin were measured by specific immunofluorescence techniques. Globin accumulated within a high proportion of HeLa cells for at least 25 h following injection, indicating the intracellular stability of the heterospecific globin mRNA and the fact that mRNA initially associated with more complex structures could be translated in the HeLa cytoplasm. In similar experiments, duck reticulocyte polysomes were translated in duck embryo cells but not in embryonic chick liver cells. The latter did withstand the microinjection procedure as judged by their capacity to respond normally to inducers and inhibitors of the synthesis of .alpha.-aminolevulinic acid synthetase. HeLa cells injected with duck Hb divide at normal or nearly normal rates following the injection, but their globin content diminishes, in contrast with the increasing globin content of cells receiving duck reticulocyte polysomes, mRNP particles or free mRNA.