Words similar to ttt
Example sentences for: ttt
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The mutagensis primers were as follows: for mutC, 5' GCTCGACCT CCA TTC TAA TG T GGG CAG GTG GTT TCA CAG 3'; mutK, 5' CCA TTC TAA TTG GGG C TT TTT GTT TCA CAG GGA GGA AGT 3'; mutE, 5' TTT CAC AGG GA C CTT GTT GAC AGT TCA ACT TCA AAC 3'; Xmut1, 5' CAC AGG GAG GAA GTT CTG TCA TCA ACT TCA AAC AGG 3'; Xmut2, 5' GAG GAA GTT GAC AGT CTA AGA CT A AAC AGG GTT GGC AGT 3'; Xmut3, 5' TGA CAG TTC AAC TTC GTT AGT GGT TGG CAG TGA CAC 3'; Ymut1, 5' TGG CAG TGA CAC AGG TGT ACT CTG CCT GCT TCC CCT 3'; and Ymut2, 5' ACA CAG GCC CAC ACT G AT GCG TTC CCC TTT CCA CCC CTG 3'.
The consensus codon is given as ttt (Phe; Table 1, column 1).
To sequence the junction fragments from both strands, we performed PCR and sequencing with primers 774F and reverse primer E43+355 (5'-CAA AGT CTC TGT CTG GAA GAG AAA TGA TTT TGA-3') for FB774, and with forward primer E44 -343 (5'-CCT TAG CTT CCC AAA GTG CTA GGA TTA-3') and 890R for FB890.
Replacement of the endogenous rec12 +locus with the rec12-Y98F allele was achieved by transformation and a pop-in, pop-out approach [ 61 63 ] . A 1.2 kbp Xba I- Xba I fragment of rec12 +was subcloned into pBluescript SKII(+) and subject to oligonucleotide directed mutagenesis [ 64 ] to simultaneously introduce a silent mutation generating an Eco RV site and to mutate the Y98 codon (GAG ATA TTT ATT ACA → GA G ATA T
Binding reactions were carried out at room temperature for 30 min in a total volume of 15 μl and contained 2-5 μg of nuclear extracts, 5 μl of 5X gel shift binding buffer (20% glycerol, 5 mM MgCl 2 , 2.5 mM EDTA, 2.5 mM DTT, 250 mM NaCl, 50 mM Tris-HCl, pH 7.5), 2 μg poly (dI-dC) and 3 × 10 4cpm/μl of [ 32P] labeled CREB (5'-AGA GAT TGC CTG ACG TCA GAG AGC TAG-3'), NF-κB (5'-AGT TGA GGG GAC TTT CCC AGG C-3') (Promega Gel Shift Assay Systems, Madison, WI, USA) or CCAAT/enhancer binding protein (C/EBP) (5'-TGC AGA TTG CGC AAT CTG CA-3') (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) oligonucleotides.
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