Restriction digests of the clone give the following sizes (kb): EcoRI--3.0, 1.7, 1.35, 0.74; HindIII--5.8, 0.6, 0.37; KpnI--6.7; KpnI/DraI--2.8, 2.0, 0.74, 0.6, 0.45; KpnI/BamHI--6.7. KpnI/XhoI excises the insert with some polylinker; KpnI/DraI excises only insert. Contains nucleotides 26-2850 of the published sequence. The KpnI-XhoI fragment contains the entire coding sequence and 133 bp of pRSVcat. The orientation is 5'KpnI to XhoI'3. The DNA-binding domain of the protein extends from aa 421 to aa 486 (encoded from nt 1393 to nt 1590). The ligand-binding domain of the protein extends from aa 528 to aa 777. Expresses functional receptor when transfected into COS-1 cells. Restriction digests of the clone give the following sizes (kb): HindIII--5.8, 0.3, 0.16; KpnI--6.4; KpnI/XbaI--6.4; XhoI--6.4; XbaI--uncut. Detects mRNA of 5.6, 6.1, and 7.1 kb in human fibroblast cell line HT-1080. As described, the construct contains an SV40 ori, the RSV LTR, pBR322 sequences, and an SV40 polyadenylation signal. |
