[Analysis of the formation and autonomous replication of an extrachromosomal mouse transgene].
A stable autonomously replicating shuttle transgene, pr8a was, previously isolated from Bombix mori and characterized. Autonomous replication of pr8a and its derivatives was observed in yeast cells, B. mori embryos, and transgenic mice. To continue the previous studies, transgenic mice of several generations were examined. DNA analysis revealed a course of pr8a rearrangements resulting in extrachromosomal transgene p8-2 of F2 transgenic mice. Consecutive directional elimination of vector DNA fragments was characteristic of rearrangements affecting autonomous transgenes in mouse cells. Analysis of the data obtained with transgenic mice and other organisms made it possible to identify the pr8a fragment that acts as minimal ARS in transformed yeast cells and transgenic mice. Another pr8a region was assumed to be a component of a pr8a ARS module involved in regulating autonomous replication. Inserted in two different integrative vectors, minimal ARS ensured their autonomous state in transformed yeast cells and transgenic mice. Transgenes pr8a and p8-2, along with some other well-known constructs, were considered as prototype autonomously replicating vectors suitable for studying the mechanism of autonomous replication and solving some problems of gene therapy.
Nikolaev
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Andreeva
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D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics; D003001:Cloning, Molecular; D004261:DNA Replication; D005111:Extrachromosomal Inheritance; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D010957:Plasmids; D019076:Transgenes