Restriction digest of the clone give the following sizes (kb): EcoRI--4.0, 1.55; BamHI--4.95, 0.6; HindIII--4.3, 0.8, 0.4; PstI--4.65, 0.9; PvuII--2.2, 1.8, 1.55. The insert contains the following restriction sites (approximate kb from the 5' end): BamHI--0.55; HindIII--0.42, 1.26; NdeI--1.06; NcoI--0.53; PstI--0.65; PvuII--0.70; SphI--1.42. Contains the complete coding sequence (nt 56-1042). The following primers can be used to amplify an 800 bp DNA fragment contained in the gene: upstream 5'-GCTTGAAGCTCTGTCCTGGGA-3'; downstream 5'-CCTAGGACATAGTAGGG-3'. When transfected into COS-7 cells, expresses a high-affinity receptor. Detects a 5.0 kb RNA transcript in human pituitary. Transcription from the T7 promoter gives the sense strand. |
