Divergent activity of the gonadotropin-releasing hormone receptor gene promoter among genetic lines of pigs is partially conferred by nuclear factor (NF)-kB, specificity protein (SP)1-like and GATA-4 binding sites

Table 2 Primers used to generate reporter vectors

Name	Sequence
−5118pGL3 F	5’-CAGACAATTAGATTCCAGGGC-3’
Promoter R	5’-TCCTTCCCCAACTGATGTAG-3’
μSF1pGL3 F^a	5’-AAGTACACAAAACAAGTTGCGGCCGGCTCTTTCACATTAAATATA-3’
proximal A OF	5’-GTTATGTGGAAGAGCCGGTG-3’
proximal OR	5’-CTTTATGTTTTTGGCGTCTTCC-3’
MμGATAUEpGL3F^a	5’-TTGCAGAAACCTAACCCCACTAGTAGGCACTAATCCAGTGTC-3’
CμNF-κBpGL3 F^a	5’-TTGGCTTGCAGAAACCTAGAATTCTATTTCCACTGAGAGCAA-3’
distal OF	5’-CAGAGAATGCTATTGCTCTC-3’
distal OR	5’-GTGTAAGTGTTGGAACCACATC-3’
Mμ1240pGL3 F^a	5’-CATAGCACCAAGGAAGCTATGCATACTAGTGGATGATACTGTGTGCAG-3’
proximal B OF	5’-AGGCACTAATCCAGTGTCTGC-3’
proximal OR	5’-CTTTATGTTTTTGGCGTCTTCC-3’
MμSP1pGL3 F^a	5’-ACCAAGGAAGCTTCCTCAACTAGTGGATGATACTGTGTGCAG-3’
MμNF-κBpGL3 F^a	5’-CATAGCACCAAGGAAGCTATGCATGGGCCTGGATGATACTGT-3’
MμGATA4pGL3 F^a	5’-ATTAGATTGCATACAAAGCTGCAGAAACAAATATTCATATTA-3’
proximal C OF	5’-TACTCCTCTTGATTTCTGACTC-3’
proximal OR	5’-CTTTATGTTTTTGGCGTCTTCC-3’

^aBlock replacement mutation reporter vectors were generated with outer forward (OF) or outer reverse (OR) primers as described in the Methods. The proximal OR resides in pGL3 3’ of the insert. Underlined bases indicate the new restriction enzyme digest site that replaced the transcription factor binding site in the native sequence

ISSN: 1477-7827