. Author manuscript; available in PMC: 2014 Jun 20.

Published in final edited form as: Mol Pharmacol. 2005 Aug 11;68(5):1239–1253. doi: 10.1124/mol.105.013417

TABLE 4.

pM and pVP16 (BD Biosciences, Palo Alto, CA) clones

In frame cloning of sequences into pM and VP16 generate fusion proteins of the GAL4-DNA binding domain and the viral protein 16 (VP16) activation domain, respectively. These plasmids were used in combinations to access interaction of proteins in mammalian cells. The multiple cloning sites of the plasmids are identical, therefore cloned sequences are interchangeable. The luciferase reporter employed in the mammalian 2-hybrid assays was pFR-Luc (Stratagene, La Jolla, CA).

Name	Primers	Amino Acids	Restriction Sites
CAR1	FP: GATCGAATTCGTCATGGCCAGTAGGGAAGATGAG	1–348	EcoR1/Xba1
	RP: BGH
CAR1-LBD	FP: GATCGAATTCGCCACCATGGTACTGTCGGCAGAAGCCC	80–348	EcoR1/Xba1
	RP: BGH
CAR3-	FP: GATCGAATTCGTCATGGCCAGTAGGGAAGATGAG	1–352	EcoR1/Xba1
	RP: BGH
CAR1-LBD	FP: GATCGAATTCGCCACCATGGTACTGTCGGCAGAAGCCC	80–352	EcoR1/Xba1
	RP: BGH
SRC-1 RID	FP: GATCGAATTCCCTAGCAGATTAAATATACAACCAG	570–780	EcoR1/Xba1
	RP: GATCTCTAGATCACATCTGTTCTTTCTTTTCCACTT
RXRα	FP: GATCGAATTCGCCGCCATGGACACCAAACATTTCCTG	1–462	EcoR1/Xba1
	RP: GATCTCTAGACTAAGTCATTT GGTGCGGCGC
RXRα-LBD	FP: GGAATTCGCCGCCATGGGCATGAAGCGGGAAG	198–462	EcoR1/Xba1
	RP: GATCTCTAGACTAAGTCATTT GGTGCGGCGC

FP, forward primer; RP, reverse primer.