By Dr. Max E. Gottesman, Henry J. Vogel
Offers advances in eukaryotic genetic recombination and discusses such subject matters as particular DNA integration in mammalian genomes, the manipulation of the mouse genome, genetic recombination in drosophila, caenorhabditis and yeast, and genome reorganization and genetic recombination in protozoa
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9, 177-184. Jaenisch, R. (1988). Science 240, 1468-1471. , and Jaenisch, R. (1983). Nature (London) 304, 315-320. , and Jaenisch, J. (1984). Proc. Nati. Sci. A. 81, 1504-1508. , and Breindl, M. (1986). Nature (London) 320, 365-367. , and Jaenisch, R. (1984). Cell 38, 597-607. , and Jaenisch, R. (1987). Genes Dev. 1, 366-375. Van Valen, P. (1966). /. Embryol. Exp. Morphol. 15, 119-124. , and Jaenisch, R. In preparation. , and Jaenisch, J. In preparation. Gray, D. Unpublished data. 6 Transfer of Yeast Artificial Chromosomes into Cultured Cells: A New Method for Manipulating the Mammalian Genome VASSILIS PACHNIS, LARYSA PEVNY, RODNEY ROTHSTEIN, AND FRANK COSTANTINI Department of Genetics and Development Columbia University New York, New York 10032 INTRODUCTION The ability to transfer cloned genes into mammalian cells, via methods such as calcium phosphate precipitation, protoplast fusion, electroporation, or microinjection, has been an invaluable tool for studying mechanisms of gene regulation, as well as for investigating the functions of specific gene products in cellular differentiation and physiology.
Figure 3 shows an example of such an experiment. After the first amplification of the DNA pools, a Fig. 3. Analysis of an experiment to subclone mutated ES cells after injection of the con struct pH/O-F (A) genomic DNA from 5 subclones (100 ng each) was pooled and amplified by PCR for 30 cycles to yield the indicative 3' fragment. 1 P32-labeled homeobox probe. Shown is the ethidium bromide-stained agarose gel and the autoradiogram. Notice the strong hybridization signal from the subclones 21-25.
4). The results are summarized in Table II. We injected a total of 681 MF1 and C57BI/6J blastocysts with the clone P4 and obtained 6 male chimeric animals. The overall chimerism in these males was poor and did not exceed 10%, as judged by coat color. The animals were mated with albino or C57BI/6J females. Five males were fertile, but from 138 offspring analyzed, none exhibited the black-agouti coat color. We next injected the clone A90 into MF1, C57BI/6J, and NMRI TABLE Π Injection of Clones P4 and A90 into Blastocyst from MF1, C57BI/6J and NMRI Strains« a Chimera Host Blastocyst transferred Number born M F Perinatal deaths Fertile males No.