Cloning of β2 adrenergic receptors c DNA and construction of yeast expression plasmid

This study aimed to clone pig β2AR gene,analysize its sequence and construct its recombinant yeast expression plasmid. Total RNA was extracted from pig liver, and then RT- PCR was conducted based on published pig β2AR gene sequence(AF000134). By cloning of the PCR product and identification,the c DNA fragment of pig β2AR was obtained. Bioinformatical tools were adopted to analyze the gene sequence and amino acid. The target gene was cloned to p PICZα A vector,constructing recombinant expression plasmid.The c DNA fragment of pig β2AR(KF023571.1) was 1257 bp,and encodeed 418 amino acids. The deduced protein was predicted to have a computed molecular mass of 46.73 ku. There were high similarity(more than83%) and homology in β2AR gene between pig and other species. Finally,recombinant plasmid named p PICZαA- β2AR was constructed successfully after verification by PCR, restriction enzyme analysis and sequencing. This research will serve as a base for expression of pig β2AR gene in yeast expression system and its application in multiresidue determination of β2-adrenoceptor agonists.