As the initial transposition reaction target DNA, we used a KpnI digest of the identified BAC clone (110 kb), which contained the exon 3 of the mouse Cdh22 gene in a 9 kb fragment.
Rpmar12 also display this dynamics, although it is now inactive, which indicates that the transposition suddenly stopped after the initial transposition burst, maybe due by the rapid loss of all active copies.
Although statistically significant, diversity-dependence remains moderate, and affects the transposition rate only marginally (the current transposition rate for all families is more than 85% of the estimated initial transposition rate when one copy only was present in the genome).
Although we did not clone and sequence the complete retroelement cluster in p1-ww[4Co63] p1-ww[4Co63]itely that both clusters is 4Co63 and B73 are identical, at least in their initial transposition of Eninu and their nested insertions of Ji and Opie.
"In the initial manner described..."....
The initial in vitro transposition reaction mixture (20 µl) contained 0.185 pmol (250 ng) transposon DNA (Kan/Neo-loxP-Mu), 500 ng target BAC DNA digested with an appropriate restriction enzyme, 2.46 pmol (176 ng) MuA, 25 mM Tris-HCl, pH 8.0, 0.05% (w/v) Triton X-100, 10% glycerol (v/v), 120 mM NaCl and 10 mM MgCl2.
More errors occurred in the "transposition" than in the "no transposition" condition (71.9 ± 21.3%% of all errors; x ± SD; paired t test: t = 2.905, df = 7, P = 0.023).
To check for changes in the transposition rate, we fit models in which transposition rates vary exponentially with time.
In it, the initials "A.C".
The increases in the transposition products closely paralleled the increase in the Tetr-labeled transposition products in the R+ cultures and in the R− cultures.
