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We used the 'Multisite Gateway' system (Invitrogen) for a modular transgene design from single His-GUs (supplementary Fig S1 online).
The respective transgenes carried one or three His-GUs integrated into the same landing site on chromosome 3R and were assayed as homozygous individuals.
DOI: http://dx.doi.org/10.7554/eLife.02443.007 10.7554/eLiFigure43 figuregure 3—figure supplement 1. Cell cycle progression of His C mutant embryos with two His-GUs ( 2xHis-GUs ).
We used the genuine His-GUs for histone expression instead of an artificial expression system, thereby maintaining S-phase-specific histone expression and avoiding histone overexpression.
According to the genome annotation, the histone gene complex of D. melanogaster is composed of 23 canonical histone gene repeat units (His-GUs), each bearing a gene array encoding the five canonical histones (for simplicity we refer to them as 'histones'; for details of the histone complex see http://flybase.org; Fig 1A).
Mutant embryos homozygous for Df 2L His C and carrying two or six transgene-encoded His-GUs were identified by lacking green fluorescent protein expression, selected from 22- to 24-h embryo collections, dechorionated, fixed for 10 min in 1 1 heptane/methanol and mounted in 1 1 Hoyers medium/lactic acid.
Autism is so much a part of his Gus-ness.
The next day, Reid confessed to me that once he'd got back to his room that evening and taken off his gi, a fish had fallen out of his karate pants.
There was artistry in the violence of his punches that pierced mid-air, the snap of his gi so sharp when he kicked that I thought it might start smoking.
Lethality of His C was not rescued by two or six transgene-encoded His-GUs.
The Rolling Stone Keith Richards has written a book about grandfathers, based on his relationship with his grandpa, Gus.
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