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Example sentences for: pgex-

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RGS3 protein fused to glutathione-S-transferase (RSG3-GST fusion protein) was produced in the pGEX-5x-1 system of Pharmacia as detailed previously [ 9 ] . 35S-Met labeled Gα-proteins were prepared in vitro using a transcription/translation protocol [ 9 ] . cDNAs for Gqα [ 25 ] and Gzα [ 26 ] were obtained from M. I. Simon; for Gsα (ref.
Wild type p53 or p53 bearing mutations in serines 33 or 37 were subcloned into pGEX-2T GST fusion vector (Pharmacia, NJ) and p53-GST purified as previously described [ 33].
GST-RANKL was purified as described previously [ 32 ] . Briefly, a fragment of murine RANKL cDNA was cloned inframe into the pGEX-4T vector (Amersham Pharmacia, Piscataway, USA) and expressed in BL21 bacteria (Amersham Pharmacia) induced with 0.1 mM IPTG (Gibco) for 5 hours at 30°C.
To generate pGa14-m.nocturnin, a fragment from the m.nocturnin cDNA [ 21 ] corresponding to the entire coding sequence (with 9 amino acids upstream of the initiation codon) was excised from pGEX-3X upon Sma I and Dra I digestion, and was inserted into the pGaL4PolyII vector opened by Bam Hl, after Klenow treatment of both vector and insert.
pGST-p35-C/EBPβ was constructed by inserting the 1,739 bp EcoRI fragment from pRSETB-EFII [ 34] into EcoRI cut pGEX-4T-1 (Amersham-Pharmacia).