Abstract:
Objective: To construct recombinant expression vectors for function epi-position of androgen receptor so as to provide foundation for further investigation and analysis interaction mechanism in vitro beween LRP16 and AR.
Methods: The plasmid pCMV-AR was amplified by using PCR technique.PCR product was cloned into the pGEM-T expression vector.Clones containing the vectors were selected on LB-plus ampicillin(100 μg·mL
-1) plates and sequenced.Plasmid DNA was isolated and purification after enzymatic digestion.AR fragments were initially connected with pCDNA3.1 vector.The recombinant plasmids pcDNA3.1-ARN1,pcDNA3.1-ARN2,pcDNA3.1-ARN3,pcDNA3.1-ARC1 were extracted and confirmed by digestion with BamHⅠand XhoⅠenzymes.
Results: Sequence analysis showed the ARN1,ARN2,ARN3,ARC1 gene fragments cloned were 100% sequence homology with that of corresponding gene coding sequence in GenBank.
Conclusion: The recombinant plasmid pcDNA3.1-ARN(1-3),pcDNA3.1-ARC1 was successfully constructed,and they could be used in vitro transcription and interpretation studies beween LRP16 and AR.