Injected amphibian oocytes: a living test tube for the study of eukaryotic gene transcription? 1977

E M de Robertis, and J B Gurdon, and G A Partington, and J E Mertz, and R A Laskey

Somatic-cell nuclei injected in Xenopus laevis oocytes remain transcriptionally active for up to 4 weeks. The rate of RNA synthesis increases as the somatic-cell nuclei enlarge. The message activity of the RNA synthesized by the injected nuclei can be demonstrated, as new proteins are made a few days after injection of HeLa-cell nuclei. Several lines of evidence demonstrate that these proteins are coded for by the HeLa-cell genome and that they result from RNA synthesized within the oocytes in the course of several days. Furthermore gene expression by the somatic-cell nuclei is selective, since only a few gene products can be detected. The oocyte cytoplasm is able to reprogramme the injected nuclei in such a way that only genes of the type normally active in oocytes are expressed. We describe experiments which indicate that genes previously inactive in the somatic cells can be switched on by the oocyte cytoplasm. These experiments have involved the use of the frog somatic-cell nuclei injected into newt oocytes. In addition, we have also found that even purified DNA is transcribed when injected into the nucleus of Xenopus oocytes. The identification of the molecules presumed to regulate chromatin transcription has been hampered by the lack of a satisfactory experimental system for assaying the biological activity of purified macromolecules. Living oocytes injected with somatic-cell nuclei (or cloned DNA) may provide such a system, since these regulatory factors could be introduced by a second injection.

UI MeSH Term Description Entries
D007525 Isoelectric Focusing Electrophoresis in which a pH gradient is established in a gel medium and proteins migrate until they reach the site (or focus) at which the pH is equal to their isoelectric point. Electrofocusing,Focusing, Isoelectric
D008845 Microinjections The injection of very small amounts of fluid, often with the aid of a microscope and microsyringes. Microinjection
D009865 Oocytes Female germ cells derived from OOGONIA and termed OOCYTES when they enter MEIOSIS. The primary oocytes begin meiosis but are arrested at the diplotene state until OVULATION at PUBERTY to give rise to haploid secondary oocytes or ova (OVUM). Ovocytes,Oocyte,Ovocyte
D010063 Ovum A mature haploid female germ cell extruded from the OVARY at OVULATION. Egg,Egg, Unfertilized,Ova,Eggs, Unfertilized,Unfertilized Egg,Unfertilized Eggs
D002467 Cell Nucleus Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed) Cell Nuclei,Nuclei, Cell,Nucleus, Cell
D002843 Chromatin The material of CHROMOSOMES. It is a complex of DNA; HISTONES; and nonhistone proteins (CHROMOSOMAL PROTEINS, NON-HISTONE) found within the nucleus of a cell. Chromatins
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D005260 Female Females
D006367 HeLa Cells The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for, among other things, VIRUS CULTIVATION and PRECLINICAL DRUG EVALUATION assays. Cell, HeLa,Cells, HeLa,HeLa Cell
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

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