The cloning of human OPRMI-EXON1 and preparation of its probe / 西安交通大学学报(医学版)

ABSTRACT

Objective To clone and sequence human OPRMI-EXON1, mark it by way of nonisotope-biotin-label, and prepare its probe to study the expression and function of human OPRMI-EXON1. Methods The target gene fragment was amplified by polymerase chain reaction (PCR), and connected to the pGEM-T vector plasmid, then recombined and cloned in competent cell. After that, it was identified by cutting with restriction endonucleases and gene sequence. Finally, we marked it and prepared its probe by nonisotope-biotin-label technique. Results It was demonstrated that the target gene length (2.2kb) amplified by polymerase chain reaction had the same size with the reckoned size in theory and had the same sequence with that of NCBI database. The probe which was used to study the opioid receptor gene was successfully prepared. Conclusion The human OPRMI-EXON1 can be successfully cloned and the probe successfully prepared from the genome, which creates a favorable basis for further research of the morphine-related genes and the expression of their dependence.