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Example sentences for: hccr

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After denaturation in 6 M and renaturation in 0.187 M guanidine-HCl in HB buffer (25 mM Hepes pH 7.2, 5 mM NaCl, 5 mM MgCl2, 1 mM DTT), the blots were saturated at 4°C in buffer H (20 mM Hepes pH 7.7, 7.75 mM KCl, 0.1 mM EDTA, 25 mM MgCl 2 , 1 mM DTT, 0.05% NP40, 1% dry milk) for 2 hours, and then incubated for 2 hours at 4°C with 50 μl of 35S-methionine-labeled in vitro-translated hCCR4 or luciferase protein (synthesized using a reticulocyte lysate-coupled transcription/translation kit [Promega]).
4B, co-expression of pGal4-hCCR4 with either pVP16-hCAF1 or pVP16-hPOP2 elicited a significant increase (> 35 fold) in pG4-TK-Luc expression, thus demonstrating association between hCCR4 and either hCAF1 or hPOP2.
To generate pGaL4-hCCR4, a fragment corresponding to the entire hCCR4 coding sequence (with the exception of the initiation codon) was obtained upon digestion of the KIAA1194 cDNA (accession number AB033020, gift from the Kazusa Research Institute, putative start codon -Met 19-positioned by homology with mCCR4) by Sph I and Msc I (nt 423 to 2172 in KIAA1194), and was inserted in-frame into the pGaL4PolyII vector opened by Xho I, after Klenow treatment of both vector and insert.
Interestingly, we found that hCCR4 also binds to the second human yPOP2 paralog, hCAF1.
The human protein (named hCCR4) is putatively encoded by the KIAA1194 full-length cDNA ( [ 27 ] , accession number AB033020).