Inducible expression of a cloned heat shock fusion gene in sea urchin embryos
-
1.
California Institute of Technology
Abstract
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.
Additional Information
© 1984 by the National Academy of Sciences. Contributed by Roy J. Britten, August 3, 1984. We are much indebted to Dr. Igor Dawid for supplying the plasmid hsp-cat1. The CAT enzyme preparation was the gift of Drs. G. Smith and M. Summers. We thank Drs. N. Davidson, E. Rothenberg, and B. Wold for thoughtful comments on the manuscript. This work was supported by the National Institute of Child Health and Human Development (HD05753). A.P.M. was supported by American Cancer Society California Division Fellowship J-33-82, and T.J.N., by National Institutes of Health Training Grant GM07616. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.Acknowledgement
We are much indebted to Dr. Igor Dawid for supplying the plas-mid hsp-catl. The CAT enzyme preparation was the gift of Drs. G.Smith and M. Summers. We thank Drs. N. Davidson, E. Rothenberg, and B. Wold for thoughtful comments on the manuscript. Thiswork was supported by the National Institute of Child Health andHuman Development (HD05753). A.P.M. was supported by American Cancer Society California Division Fellowship J-33-82, andT.J.N., by National Institutes of Health Training Grant GM07616.
Files
| Name | Size | Download all |
|---|---|---|
|
md5:1dc4a307512084445082c0d7c68a1cda
|
1.4 MB | Preview Download |
Additional details
- NIH
- HD-05753
- American Cancer Society, California Division
- J-33-82
- NIH
- GM07616
- Created
-
2007-10-23Created from EPrint's datestamp field
- Updated
-
2019-10-02Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering (BBE)
- Publication Status
- Published