I would like to produce a truncated form of the GPCR we are investigating at the lab, i.e. a receptor lacking a part of the intracellular C-tail, with the Transformer site-directed mutagenesis kit of Clontech (in vitro site-directed mutagenesis into a target gene or region cloned into a double-stranded plasmi). Which of the two posibilities that come to my mind is the better?
1) simply convert an aminoacid codon in a stop codon.
2) delete the part of the receptor that we don't want (about 15 aminoacid) , maintaining the original stop codon in the plasmid.
The first one seems to me so simple... Does the new construct will effectively result in a truncated protein? In case this is considered the best approach, do the stop codons are equivalent or one is better that the others?
Thank you in advance for your assistance and patience...Val