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Gene Transfer by Direct Pronuclei Microinjection

  • Albert W. Tam
Part of the Basic Life Sciences book series

Abstract

The technique of micromanipulating and injecting a mouse egg was first described in 1966 by T.P. Lin (8,9). With the advent of recombinant DNA technology, it has become possible to transform cultured cells by modifying this technique to microinject directly into the cell nuclei-specific cloned gene sequences (1,4). This approach is amenable only to established cell lines in culture, and so its general usefulness in the analysis of tissue-specific gene expression has been found to be somewhat limited. Subse-quently, transgenic animals have been produced by introduction of foreign genes at the pronuclear stage of fertilized, one-cell zygotes. Most of the successes have been with mouse eggs (3,5,6,11,12), and only recently has the successful production of transgenic animals been extended to the rabbit, pig, and sheep (7). This technique has become a powerful tool for studying gene regulation and physiological functions of gene products in a normal host environment. This chapter summarizes the current methodology used for mouse eggs in gene transfer experiments by direct pronuclear microinjection.

Keywords

Thymidine Kinase Cumulus Cell Male Pronucleus Injection Chamber Female Pronucleus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1986

Authors and Affiliations

  • Albert W. Tam
    • 1
  1. 1.Department of Molecular BiologyGenentech, Inc.South San FranciscoUSA

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