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Page 1 \ ?^p 6r.1 CELL CYCLE CONTROL OF HUMAN H4 ...

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a2t2 F: call cyds contol d u4 G@ f,wiPl/ol<br />

Fig,7. sch6hatic dlagnm showlng the ctonlng procedore tor <strong>H4</strong><br />

fiutant promoler conslrucis. (A) Ths top $rsnd oi lhe t€st<br />

oiigo and bottorn strand ol rhe adaPtor oligo w€€ each<br />

phosphorylatsd and ann€alBd ,rte trayod ends w8re frll€d in<br />

wth Krenow and digested wirh Psl L (a) pFo10€ sas d6av€d<br />

wirh Pst | / Nae I and lh6 oligonud€otide casselt€ trom (A) was<br />

rigated into ih€ Pst | / Na€ | sires ot <strong>H4</strong> veclor puc19 which<br />

r6sults in rhe inlermedialr dorc A. (C) pFOl0€ CAT was c{€av€d<br />

with Pst I ro isolar€ lhe caT tragment and ligaled into linearlz€d<br />

inbem€diate clono A, rssulting in site ll <strong>H4</strong> mutant promoEr CAI<br />

conslrucr (o).<br />

32

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