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Molecular cloning and characterization of a novel beta-adrenergic receptor.

X.H. Chen,T. K. Harden,Robert A. Nicholas-1994-10-01-Journal of Biological Chemistry
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TL;DRAbstract

In an attempt to isolate new G protein-coupled receptors from turkey erythrocytes, reverse transcribed polymerase chain reaction was performed on fetal turkey blood RNA using degenerate primers based on conserved sequences present in seven transmembrane receptors. An open reading frame in one of the clones, designated 4C (497 base pairs), displayed approximately 50-60% identity to all of the previously cloned beta-adrenergic receptors (beta-ARs). A lambda-DASH turkey genomic library was screened with a probe generated from the partial 4C cDNA, and the gene encoding this receptor was localized to a 3.5-kilobase pair HindIII fragment. Ribonuclease protection analysis of turkey lung mRNA indicated that the 3' end of the coding sequence of the 4C gene, like beta 3-AR, was interrupted by an intron. To obtain the cDNA sequence of 4C, RNA-polymerase chain reaction was performed using primers complementary to regions identified by ribonuclease protection analysis to be present in 4C mRNA. Comp

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In an attempt to isolate new G protein-coupled receptors from turkey erythrocytes, reverse transcribed polymerase chain reaction was performed on fetal turkey blood RNA using degenerate primers based on conserved sequences present in seven transmembrane receptors. An open reading frame in one of the clones, designated 4C (497 base pairs), displayed approximately 50-60% identity to all of the previously cloned beta-adrenergic receptors (beta-ARs). A lambda-DASH turkey genomic library was screened with a probe generated from the partial 4C cDNA, and the gene encoding this receptor was localized to a 3.5-kilobase pair HindIII fragment. Ribonuclease protection analysis of turkey lung mRNA indicated that the 3' end of the coding sequence of the 4C gene, like beta 3-AR, was interrupted by an intron. To obtain the cDNA sequence of 4C, RNA-polymerase chain reaction was performed using primers complementary to regions identified by ribonuclease protection analysis to be present in 4C mRNA. Comp

Keywords

Molecular biologyBiologyComplementary DNAExonGeneUntranslated regionPeptide sequenceSequence analysis

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