Construction of recombinant eukaryotic expression plasmid of human thyrotropin receptor extracellular domain encapsulated with cationic liposomes

Abstract

AIM: To construct recombination eukaryotic expression plasmid of human thyrotropin receptor extracellular domain encapsulated with cationic liposomes. METHODS:We amplified the target gene of shuttle vector PHMCMVTSHR289, conjugated the target gene and eukaryotic expression plasmid pcDNA3.1+, and accredited whether pcDNA3.1+/TSHR289 was connected or not by enzymatic digestion and sequencing. Cationic liposomes encapsulated the recombination plasmid pcDNA3.1+/TSHR289.RESULTS: Recombination plasmid pcDNA3.1+/TSHR289 digested with enzyme HindIII and the fragment through 0.8% gel electrophoresis showed 512bp strip. Recombination plasmid pcDNA3.1+/TSHR289 were found synonymous mutation through forward(AAC to AAT)and reverse sequencing(GCG to GCT). The volume ratio of cationic liposomes and recombinant plasmid was 3:1. CONCLUSION: It is successful to construct the recombination plasmid pcDNA3.1+/TSHR289 by accredit it through enzymatic digestion and sequencing.

Keywords

• human thyrotropin receptor extracellular domain gene
• shuttle vector PHMCMVTSHR289
• eukaryotic expression plasmid pcDNA3.1+
• recombination plasmid pcDNA3.1+/TSHR289
• cationic liposomes