Nathalie Troffer-Charlier, Vincent Cura, Pierre Hassenboehler, Dino Moras, Jean Cavarelli
نویسندگان
چکیده
Cloning of CARM128-140 The N terminal part of mouse CARM1 (amino-acids 28-140) was PCR amplified from the original GST-CARM1 construct (Chen et al, 1999) and cloned in the expression vector pGGWA (Busso et al, 2005) using the Gateway system (Invitrogen). The primers used for cloning were 5’-GGG GAC AAG TTT GTA CAA AAA AGC AGG CTT CCT GGT GCC ACG CGG TTC TCA TAT GGC TAC AGT GTC TGT GTT CCC-3’ with a NdeI site and a thrombin cleavage site and 5’-GGG GAC CAC TTT GTA CAA GAA AGC TGG GTC AGA TCT TCA CTC ACT GAA CAC AGA GCG C-3’ with a BglII site. The DNA sequence of the amplified domain was entirely verified by sequencing. The protein is expressed as a GST fusion. After thrombin cleavage, the peptide GSHM remains at the N terminal end of the protein and is effectively seen in the final electron density map.
منابع مشابه
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