Table 1. IL-6 promoter constructs... | American Society of Hematology

Table 1.

IL-6 promoter constructs

Construct name	Mutation	Purpose of mutation	Primers (restriction enzyme site)^*,^†
pIL6-1200/SEAP	None	N/A	See text
pIL6-327/SEAP	Deletion of −1200 to −327	Isolate effects of all the known positive regulatory elements located between −327 and +1.	F = 5′-GGAAGATCTCTTCGTGCATGACTTCAGCTTTACT-3′ (BglII) R = 5′-CGGGAATTCAGGGCAGAATGAGCCTCAGAGACAT3-3′ (EcoRI)
pIL6-327-AP1mut/ SEAP	Point mutation of an AP1 site	Determine if AP1 site is necessary for LANA-mediated induction of IL-6 expression	F = 5′-TGCAATCAGATCTATGCCAAGTGCTGCAGCACTAATA-3′ (BglII) R = 5′-CGGGAATTCAGGGCAGAATGAGCCTCAGAGACAT3-3′ (EcoRI)
pIL6-225/SEAP	Deletion of −327 to −225	Deletion of AP1 and ETS sites	Generated from pIL6-327/SEAP by digestion with NheI followed by religation.

Construct name	Mutation	Purpose of mutation	Primers (restriction enzyme site)^*,^†
pIL6-1200/SEAP	None	N/A	See text
pIL6-327/SEAP	Deletion of −1200 to −327	Isolate effects of all the known positive regulatory elements located between −327 and +1.	F = 5′-GGAAGATCTCTTCGTGCATGACTTCAGCTTTACT-3′ (BglII) R = 5′-CGGGAATTCAGGGCAGAATGAGCCTCAGAGACAT3-3′ (EcoRI)
pIL6-327-AP1mut/ SEAP	Point mutation of an AP1 site	Determine if AP1 site is necessary for LANA-mediated induction of IL-6 expression	F = 5′-TGCAATCAGATCTATGCCAAGTGCTGCAGCACTAATA-3′ (BglII) R = 5′-CGGGAATTCAGGGCAGAATGAGCCTCAGAGACAT3-3′ (EcoRI)
pIL6-225/SEAP	Deletion of −327 to −225	Deletion of AP1 and ETS sites	Generated from pIL6-327/SEAP by digestion with NheI followed by religation.

Template for PCR amplification was pIL6-1200/SEAP, unless otherwise stated.

PCR amplification products were cloned intoBglII/EcoRI sites in pSEAP-Basic, unless otherwise stated.