TABLE 1.

Effect of naturally occurring and randomly designed promoter mutations on HBV replication^a

Construct	Mutation		Replicationc
	Nucleotide position(s)	Position(s)b in mutated core promoter sequence
Mutants
MT5/6	1768, 1770	TAAAGGTTTATGTATT	15
MT8	1764, 1766	TAATGATCTTTGTATT	1.5–2
MT9	1770	TAAAGGTCTGTGTATT	No change
MT10	1768, 1770	TAAAGGTATGTGTATT	No change
MT11	1768, 1770	TAAAGGTGTATGTATT	No change
MT12	1768	TAAAGGTATTTGTATT	No change
MT13	1768, 1770	TAAAGGTATATGTATT	No change
MT14	1772	TAAAGGTCTTTATATT	No change
WT adw		TAAAGGTCTTTGTATT	1

^aVarious mutations were introduced into the replication-competent construct adwR9 by oligonucleotide-directed mutagenesis. Four days after transfection of wild-type (WT) and mutant constructs into HuH-7 cells, viral replication was analyzed by Southern blotting of core particle-associated viral replicative intermediates as shown in Fig. 8A. The amount of viral replicative intermediates was quantified with a phosphor imager. Core promoter nucleotide positions are numbered according to reference 32 and MT8, represents mutations described in reference 34. 

^bUnderlined. 

^cShown as fold increase compared to replication of wild-type HBV adw (replicative intermediates of wild-type adw construct = 1).