Words similar to nfatp

Example sentences for: nfatp

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First, pBS-NFATp(391-583) was created using single-stranded pBS-KS +-NFATp(NdeI) for site-directed mutagenesis with two oligonucleotides: a unique in-frame ATG codon contained in an Nde I site was created in place of the codon for amino acid 390 using an oligonucleotide of sequence 5'-CATCTGCAGCATCCATATGACTGCATCCCTC-3' and a stop codon was created in the NFATp cDNA after amino acid 583 using an oligonucleotide of sequence 5'-GTTGAAAGACAATAAACAGAATTCTGCCTGGTCTATG-3'.
In this study, however, transcriptional activation by NFATp was not dependent on the identified transactivation domains.
Recombinant HA-tagged NFATp proteins (approximately 500 ng) were incubated with calcineurin (500 ng, Sigma) and calmodulin (200 ng, Sigma) in 21 μl of buffer that had the following final concentrations of components: 7 mM Tris-HCl, 5 mM Hepes (pH 7.6), 90 mM NaCl, 15 mM betamercaptoethanol, 0.3 mM DTT, 1 mM EDTA, 30% (v/v) glycerol, 3 mM MgCl 2 , and 1.5 mM MnCl 2 . Control reactions were performed with NFATp proteins in the absence of added calcineurin and calmodulin.
The PCR product was digested with Xba I and Mro I and ligated into pBS-KS +-NFATp that had been digested with Xba I and Mro I. pVL1392-HAX-NFATp(1-722) was made by subcloning a Nde I - Msc I fragment from pBS-KS +-NFATp(NdeI) into the Nde I and Sma I sites of pVL1392-HAX.
Thus, the N-terminal region of NFATp (amino acids 1-330) was required to observe a phosphorylation-dependent decrease in migration of the recombinant HA-NFATp.