Sir John Gurdon's famous frog cloning experiments of the 1960s and ’70s answered a question that had been hanging over cell biologists since before the turn of the century: are the cells of an adult organism genetically identical to the fertilized egg from which they are derived? The development of the egg tests the capacity of the genes in the...The transplantation of nuclei from differentiated or determined somatic First we consider the mechanism by which nucleosomes are formed from DNA and histones. Experiments demonstrate that high molecular weight RNA synthesized by these mutants attaches to polysomes active in protein synthesis.The early effects are described of injecting nuclei from embryonic, larval, and adult cells of Xenopus laevis into unfertilized eggs of the same species. We do not accept anonymous comments.
Several lin...This chapter describes the methods for nuclear transplantation in amphibia.
Please include your email address; the address will not be displayed in the posted comment. Search for other works by this author on: The synthesis, uptake, and release of RNA and proteins by injected nuclei have been investigated by autoradiography. Towards the end of cleavage a m...The injection of purified DNA into unfertilized frogs' eggs stimulates a 20-fold or greater increase of [3H]dT incorporation into DNA. clearly recognizable change of gene activity. Full length 5S RNA of the correct sequence is transcribed from the injected DNA, and the newly synthesized transcripts hybridize predominantly with the coding strand of 5S DNA. The effect of ribosome “factors” (materials released from polysomes by 0.5 or 1.0 M KCl) on the translation of messenger RNA has been tested by injecting factors and mRNA into living frog oocytes. Two main regions were studied: the yolky endoderm plus ventral mesoderm, and the axial region containing functional myotomes, nerve c...Dr Gurdon discusses the introduction of purified macromolecules, such as Single-stranded DNA and linear double-stranded DNA were degraded by both nucleus and cytoplasm.
He interpreted his discovery that the transfer of specialized intestine nuclei into enucleated frog oocytes could give rise to all the tissues of swimming tadpoles as evidence of the remarkable plasticity of which nuclei, even from highly differentiated cell types, were capable (Gurdon et al., 1958). Of the injected eggs, 30% survived to the swimming tadpole stage, and 15% became frogs. acti...In Xenopus laevis, nucleolar mutants which form a partial nucleolus have
Briggs and King in the early ’50s suggested that cells' genetic material is irreversibly altered as they begin to differentiate. The oocytes are defolliculated and enucleated manually and allowed to heal in a potassium phosphate buffer. Apparen...Single and multiple repeating units of three types of Xenopus 5S DNA recombined with the plasmid pMB9 serve as templates for the accurate synthesis of 5S RNA after their injection into Xenopus laevis oocyte nuclei.
As an example of the way in which the technique may be used, a study of the role of deoxynucleotide triphosphates in regulating DNA synthesis is described.
Ideally we'd like to have a small research institute.” They said, “That's good, but we can't pay for all that.” But by good fortune, the Wellcome Trust was willing to collaborate with CRC to fund an institute. But Gurdon showed that this was not the case. Nuclei were transplanted into enucleated eggs which contained [32P]phosphate, and the resulting nuclear transplant-embryos showed a pattern of RNA...A diversity of results has been obtained from nuclear transfer experiments in Amphibia since the time when a satisfactory technique was first developed for transplanting living cell nuclei into enucleated eggs.
Three cloned fragments of...The intracellular distribution and activity of enzymes which act on DNA in oocytes of Xenopus laevis have been investigated by injecting conformational variants of SV40 DNA into the oocyte's nucleus or cytoplasm.
The director at the time was Jim Smith, a very fine director, but there were so many Smiths in the world that it didn't seem a very helpful name. This paradox is sometimes explained by assuming that the nucleuses do not have access to DNA, from which they may be separated by the nuclear membrane. For this reason they are particularly well suited for microinjection experiments.
in about equal amounts of α and β globin synthesis in a Nuclear proteins extracted from other cells enter the injected nuclei. They were identical in size to the polypeptides that appeared in ascites cells after infection with this virus. Sephadex and carboxymethyl cellulose chromatography was used to show that rabbit or mouse globin was synthesized continuously by embryos and tadpoles up to the stage when feeding co...Fertilised eggs of Xenopus laevis were injected with rabbit or mouse globin mRNA and allowed to develop. It might be that by altering the histone state of genes that are the targets of the Yamanaka factors, we might make those genes more receptive and thus increase reprogramming efficiency. The reasons for developing the technique of nuclear transplantation are to test the genetic content of somatic cells and to analyze interactions between nucleus and cytoplasm.
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