Premium
Molecular cloning of a human thyrotropin receptor cDNA fragment
Author(s) -
Huang Guo Cai,
Page Martin J.,
Roberts Andrew J.,
Malik Afshan N.,
Spence Hugh,
McGregor Alan M.,
Banga J.Paul
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)80246-f
Subject(s) - thyrotropin receptor , complementary dna , transmembrane domain , biology , microbiology and biotechnology , nucleic acid sequence , receptor , expression cloning , peptide sequence , oligonucleotide , cloning (programming) , sequence analysis , molecular cloning , genetics , autoantibody , antibody , gene , computer science , programming language
Autoantibodies to the thyrotropin (TSH) hormone receptor (TSH‐R) are present in the sera of patients with thyroid autoimmune disease which are pathogenetic leading to hyperthyroidism of Graves' disease. Considerable interest has been focused on the cloning of the human TSH‐R, which has until very recently, proven exceedingly difficult due to the very low receptor level expression on thyroid cells. We have used polymerase chain reaction and highly degenerate, inosine containing oligonucleotides derived from sequence alignments of the transmembrane regions 2 and 7 of a number of G‐binding protein receptors including the lutropin/choriogonadotropin (LH/CG) receptors to amplify various cDNAs from human thyroid cDNA. Sequencing analysis of 27 different clones revealed that they fall into eight different groups. The very recent publication of the complete nucleotide sequence of the human TSH‐R revealed that one of the groups (GT1) containing seven clones which had been sequenced belong to the human TSH‐receptor. The sequence of all 7 GT1 clones was identical and in complete concordance with transmembrane regions 2 and 7 of the published TSH‐R sequence. Our results show that by designing oligonucleotides to common transmembrane regions of G‐binding proteins where the primers are biased in their sequence to the LH/CG receptors it is possible to amplify the TSH‐R receptor sequence.