Words similar to pcmv

Example sentences for: pcmv

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Oligonucleotide primers for the PCR reaction were based on the published sequence of murine Rab24 [ 24 ] . Kpn1 and BamH1 restriction sites were added to the 5' and 3' primers respectively, to facilitate cloning of the PCR product in frame with a 5' sequence cassette encoding a myc epitope (EQKLISEEDL), previously introduced into the pCMV5 expression vector [ 78 ] . The Rab24 cDNA was altered by overlap-extension PCR [ 79 ] , using appropriate mutator oligonucleotides and Pfu DNA polymerase (Stratagene Inc., La Jolla, CA), so as to introduce amino acid substitutions at D123 (D123I), T120 (T120A), S22 (S22N), S67 (S67Q), and a deletion of residues 127 through 130 (ΔEDRR).
Human β 1 -pCMV5 and bovine γ 2 -pcDNAI have been described [ 35 ] .
The cDNAs encoding Rab24(wt) and Rab24(D123I) were subcloned into a pCMV5 vector that had been modified by PCR to encode an in-frame polyhistidine tag (His 6 ) at the amino-terminus of the Rab protein.
HEK 293 cells grown in 10 cm diameter dishes were transfected with either pCMV5- His 6 Rab24 or pCMV5- His 6 Rab24(D123I) as described earlier.
The mammalian expression constructs of Smad1 and its deletion mutant Smad1C (271-465) were made by subcloning Smad1 into two modified pCMV6 vectors, which have either the Flag epitope (F-pCMV6) or the T7 epitope (T7-pCMV6) placed upstream of an array of new multiple cloning sites.