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Generation of an artificial ring chromosome in A rabidopsis by C re/ L ox P ‐mediated recombination
Author(s) -
Murata Minoru,
Shibata Fukashi,
Hironaka Akiko,
Kashihara Kazunari,
Fujimoto Satoru,
Yokota Etsuko,
Nagaki Kiyotaka
Publication year - 2013
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.12128
Subject(s) - human artificial chromosome , eukaryotic chromosome fine structure , centromere , chromosome , telomere , biology , genetics , yeast artificial chromosome , dna , gene , gene mapping
Summary A eukaryotic chromosome consists of a centromere, two telomeres and a number of replication origins, and ‘artificial chromosomes’ may be created in yeast and mammals when these three elements are artificially joined and introduced into cells. Plant artificial chromosomes ( PAC s) have been suggested as new vectors for the development of new crops and as tools for basic research on chromosomes. However, indisputable PAC formation has not yet been confirmed. Here, we present a method for generating PAC s in the model plant A rabidopsis thaliana using the C re/ L ox P and A ctivator/ D issociation element systems. The successfully generated PAC , designated At ARC 1 ( A. thaliana artificial ring chromosome 1), originated from a centromeric edge of the long arm of chromosome 2, but its size (2.85 Mb) is much smaller than that of the original chromosome (26.3 Mb). Although A t ARC 1 contains only a short centromere domain consisting of 180 bp repeats approximately 250 kb in length, compared with the 3 Mb domain on the original chromosome 2, centromere‐specific histone H3 ( HTR 12) was detected on the centromeric region. This result supported the observed stability of the PAC during mitosis in the absence of selection, and transmission of the PAC to the next generation through meiosis. Because A t ARC 1 contains a unique L ox P site driven by the C a MV 35 S promoter, it is possible to introduce a selectable marker and desired transgenes into A t ARC 1 at the L ox P site using C re recombinase. Therefore, A t ARC 1 meets the criteria for a PAC and is a promising vector.