6.3.4.2	-	
6.3.4.2	a carboxyl-terminal FLAG sequence is introduced after insertion of human CTPS1 into pCDNA 4 myc/his, expression by transient transfection of human embryonic kidney (HEK) 293 cells	
6.3.4.2	cDNA encoding the synthetase domain of human CTPS (isoform 1) is subcloned into the pNIC-Bsa4 vector, the resulting construct codes for a protein with an N-terminal hexahistidine tag with an integrated TEV protease-cleavage site, the entire plasmid is transformed in Escherichia coli BL-21	
6.3.4.2	cloned in Escherichia coli as a histidine-tagged fusion protein	
6.3.4.2	cloned in Escherichia coli BL-21 as an N-terminal 6xHis-tag fusion protein	
6.3.4.2	expressed in Escherichia coli BL21(DE3) cells	
6.3.4.2	expression in a ura7ura8 double mutant that lacks CTP synthetase activity	
6.3.4.2	expression in Escherichia coli	
6.3.4.2	expression of the human type II CTP synthetase gene in the CTP synthetase-deficient mutant Escherichia coli JF618 completely abolishes the need for exogenous cytidine for their survival	
6.3.4.2	expression of the mutant enzyme D149E in Escherichia coli