Inducible expression of a cloned heat shock fusion gene in sea urchin embryos.
Inducible expression of a cloned heat shock fusion gene in sea urchin embryos.
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克隆的热休克融合基因在海胆胚胎中的诱导表达。
DOI：
10.1073/pnas.81.23.7490
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发表时间：
1984
影响因子：
11.1
通讯作者：
Davidson,EH
中科院分区：
文献类型：
--
作者：
McMahon,AP;Novak,TJ;Britten,RJ;Davidson,EH
融合基因构建体，其中细菌氯霉素乙酰转移酶（CAT;乙酰辅酶A：氯霉素3- 0-乙酰转移酶，EC 2.3.1.28）的编码序列置于果蝇基因的调控区的控制下，所述果蝇基因编码70千道尔顿热休克蛋白[Di Nocera，P. P.等人（1983）Proc. Acad. Sci. USA 80，7095-7098]，显微注射到未受精的海胆卵的细胞质中。从注射卵发育而来的pluteus阶段胚胎暴露在高温条件下，我们发现这会引起内源性海胆热休克反应。这些胚胎表达CAT基因，并且在热处理后，显示出比在正常温度下培养的对照胚胎的提取物高8-10倍的CAT酶活性。注射的DNA以高分子量串联存在，并且在发育期间扩增约100倍。扩增的序列负责所有或大部分诱导的CAT酶活性。
A fusion gene construct, in which the coding sequence for bacterial chloramphenicol acetyltransferase (CAT; acetyl-CoA: chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28) was placed under the control of the regulatory region of the Drosophila gene encoding the 70-kilodalton heat shock protein [Di Nocera, P.P. & Dawid, I.B. (1983) Proc. Natl. Acad. Sci. USA 80, 7095-7098], was microinjected into the cytoplasm of unfertilized sea urchin eggs. Pluteus-stage embryos developing from the injected eggs were exposed to high temperature conditions that we found would elicit an endogenous sea urchin heat shock response. These embryos express the gene for CAT and, after heat treatment, display 8-10 times more CAT enzyme activity than do extracts from control embryos cultured at normal temperatures. The injected DNA is present in high molecular weight concatenates and, during development, is amplified about 100-fold. Amplified sequences are responsible for all or most of the induced CAT enzyme activity.