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Generation of a human X‐derived minichromosome using telomere‐associated chromosome fragmentation.
Author(s) -
Farr C. J.,
Bayne R. A.,
Kipling D.,
Mills W.,
Critcher R.,
Cooke H. J.
Publication year - 1995
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1995.tb00228.x
Subject(s) - minichromosome , biology , telomere , chromosome 22 , centromere , minichromosome maintenance , chromosome , genetics , chromosome 19 , microbiology and biotechnology , origin of replication , dna , gene , dna replication
A linear mammalian artificial chromosome vector will require at least three functional elements: a centromere, two telomeres and replication origins. One route to generate such a vector is by the fragmentation of an existing chromosome. We have previously described the use of cloned telomeric DNA to generate and stably rescue truncated derivatives of a human X chromosome in a somatic cell hybrid. Further rounds of telomere‐associated chromosome fragmentation have now been used to engineer a human X‐derived minichromosome. This minichromosome is estimated to be < 10 Mb in size. In situ hybridization and molecular analysis reveal that the minichromosome has a linear structure, with two introduced telomere constructs flanking a 2.5 Mb alpha‐satellite array. The highly truncated chromosome also retains some chromosome‐specific DNA, originating from Xp11.21. There is no significant change in the mitotic stability of the minichromosome as compared with the X chromosome from which it was derived.