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Updated: Aug 23, 2026
Functional Cloning Using a Xenopus Oocyte Expression System
Published on: January 30, 2016
[Cloning of the complete coding sequence of mouse oxytocin receptor gene and its eukaryotic expression]
Liang Jin1, Wei-lin Jin, Gong Ju
1Institute of Neuroscience, Fourth Military Medical University, Xi'an 710032, China. jin173@hotmail.com
Aim:
To clone the complete coding sequence of mouse oxytocin receptor (mOTR) gene and to express it in mammalian cells.
Methods:
Overlapping carboxyl end and amino end cDNAs of mOTR gene were acquired respectively by RT-PCR method and subcloned into PMD 18-T vectors. Through adding the digestion sites, double digestions and ligation, the 2 fragments were linked to form the complete coding sequence which was inserted into the eukaryotic expression vector pEGFP-N3, and the recombinant pEGFP-mOTR plasmid was transfected into COS-7 cells for eukaryotic expression by liposome-mediated gene transfer method.
Results:
The carboxyl end and amino end cDNAs of mOTR gene were successfully subcloned into PMD 18-T vectors. The complete coding sequence was obtained and cloned into the pEGFP-N3 vector. The recombinant pEGFP-mOTR plasmid was transiently expressed in COS-7 cells successfully.
Conclusion:
The recombinant eukaryotic expression vector pEGFP-N3-mOTR was constructed successfully and transiently expressed in COS-7 cells, which lays the foundation for further functional studies of mOTR.
