Open AccessAnalysis of the Vicia faba Genome by Use of Restriction Endonucleases
Author(s) -
KAINA Bernd,
HINZ Rainer
Publication year - 1979
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1979.tb13025.x
Subject(s) - restriction enzyme , ecori , dna , hindiii , vicia faba , genome , agarose , restriction fragment , base pair , chemistry , gc content , microbiology and biotechnology , bovine genome , restriction fragment length polymorphism , biology , genetics , gene , polymerase chain reaction , botany
DNA of the broad bean, Vicia faba , was cleaved by the restriction endonucleases endoR · Eco RI, endoR · Hin dIII, endoR · Hin cII, endoR · Bam I, and endoR · Bsp RI. Separation in agarose gels of the resulting fragments revealed, in addition to the bulk DNA, an enzyme‐specific pattern of bands composed of restriction fragments of 300 to more than 30000 base pairs in length. Bulk DNA was characterized by an unusual size distribution which significantly deviated from that expected according to the random fragmentation theory. It is argued that the observed distribution is due to the high proportion of repetitive DNA within this species (∼ 75%). In all digests, a class of high‐molecular‐weight restriction fragments of more than 30000 base pairs in length was observed which comprised 5–8% of the genome. It showed hybridization with highly repetitive DNA ( c 0 t 2 × 10 −2 M · s) and included a fraction (2–3% of the genome) highly resistant to the activity of all the enzymes tested. The buoyant density in CsCl of this resistant DNA was not different from that of the total DNA (36% dG + dC). In endoR · Eco RI digests, the high‐molecular‐weight fragment class contained, in addition to the resistant DNA, a fraction of relatively high buoyant density (calculated dG + dC content: 61%) containing cleavage sites for the other enzymes used.