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The second investigated URR bears a functional Cre1 double site [ 23].
Therefore, the following modifications were introduced (protein sequences are indicated by UNIPROT accession numbers): The first phosphorylation site of P02646 is actually a double site that lies at positions 22/23 instead of position 20 [ 59, 60].
Because KCTD15 has two predicted binding sites in its 3′-UTR, we performed molecular cloning for single site mutations to obtain vectors named MUT1-KCTD15, MUT2-KCTD15, and double site mutation vectors named MUT1,2-KCTD15 MUT1,2-KCTD15
Given the possibility of different but highly related HPV E2C domains, we compared binding of E2C domains from HPV11, HPV16, and HPV18 (E2C-16, E2C-18, and E2C-18) to individual single sites and the double site from the HPV16 genome.
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Due to the pronounced chemical shift changes in purified MEKK3-PB1-I57tfmF MEKK3-PB1-I57tfmF MEKK3-PB1-I57tfmFe two sites were selected for double site-specific landling.
The double site-directed H306Q-H310E mutageneshowshow that G. graminis LOX activity is not causing the low yield of the G. graminis LOX.
Open image in new window Figure 2 One dimensional 19 F NMR spectra of MEKK3-PB1 or MEK5-PB1 in crude lysates with double site-specific tfmF incorporation.
To test whether the activity of the G. graminis LOX interferes with the correct synthesis and secretion of the protein a double site-directed H306Q-H310E mutagenesis was performed resulting in inactive A. nidulans GG-LOX mutants.
In contrast, the Db model derived from animals with double site-directed mutations at amino acid residues 315 and 322 of the PyV-mT is decoupled from the PI3-K pathway.
We also used double sites mutated probe as negative control, which exhibited diminished DNA-protein interaction band (lane 4).
No evidence of recombination was seen in 92 meiotic events, which would be indicated by a chromosome with either no NUMT or a doubled site.
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