Table 2. Summary of reagents used for obtaining constructs of mutant derivatives of C5 protein in pFHC1008′.

Name of construct	Mutations introduced	Primers used for PCR	PCR template	Enzymes used for cloning
Group Ia
pFHC1008′-Sn49	R46H	-	-	EcoRI and HindIII
pFHC1008′-Sn32	R57A	-	-	EcoRI and HindIII
pFHC1008′-Sn9	R62A	-	-	EcoRI and HindIII
pFHC1008′-Sn24	K66A	-	-	EcoRI and HindIII
pFHC1008′-Sn25	R67A	-	-	EcoRI and HindIII
Group II
pFHC1008′-Sn7	F18A	SDMO7 and rHindIII	pFHC1008′	HpaI and HindIII
pFHC1008′-Sn16	F22A	SDMO16 and rHindIII	pFHC1008′	HpaI and HindIII
pFHC1008′-Sn716	F18A/F22A	SDMO716 and rHindIII	pFHC1008′	HpaI and HindIII
Group III
pFHC1008′-Sn33	R70A	SDMO33 and fEcoRI	pFHC1008′	EcoRI and HindIII
pFHC1008′-Sn925	R62A/R67A	SDMO925 and fEcoRI	pFHC1008′	EcoRI and HindIII
pFHC1008′-Sn933	R62A/R70A	SDMO933 and fEcoRI	pFHC1008′	EcoRI and HindIII
pFHC1008′-Sn2533	R67A/R70A	SDMO2533 and fEcoRI	pFHC1008′	EcoRI and HindIII
pFHC1008′-Sn4932	R46H/R57A	SDMOA49 and rHindIII	pFHC1008′-Sn32	EcoRI and HindIII
pFHC1008′-Sn499	R46H/R62A	SDMOA49 and rHindIII	pFHC1008′-Sn9	EcoRI and HindIII
pFHC1008′-Sn4924	R46H/K66A	SDMOA49 and rHindIII	pFHC1008′-Sn24	EcoRI and HindIII
pFHC1008′-Sn4925	R46H/R67A	SDMOA49 and rHindIII	pFHC1008′-Sn25	EcoRI and HindIII
pFHC1008′-Sn4933	R46H/R70A	SDMOA49 and rHindIII	pFHC1008′-Sn33	EcoRI and HindIII
Group IV
pFHC1008′-Sn79	F18A/R62A	SDMO7 and rHindIII	pFHC1008′-Sn9	HpaI and HindIII
pFHC1008′-Sn724	F18A/K66A	SDMO7 and rHindIII	pFHC1008′-Sn24	HpaI and HindIII
pFHC1008′-Sn725	F18A/R67A	SDMO7 and rHindIII	pFHC1008′-Sn25	HpaI and HindIII
pFHC1008′-Sn169	F22A/R62A	SDMO16 and rHindIII	pFHC1008′-Sn9	HpaI and HindIII
pFHC1008′-Sn1624	F22A/K66A	SDMO16 and rHindIII	pFHC1008′-Sn24	HpaI and HindIII
pFHC1008′-Sn1625	F22A/R67A	SDMO16 and rHindIII	pFHC1008′-Sn25	HpaI and HindIII

^aThe different strategies used for obtaining the various groups of mutant derivatives are described in the Materials and Methods.