[Cloning of the complete coding sequence of mouse oxytocin receptor gene and its eukaryotic expression].
OBJECTIVE
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.


CONCLUSIONS
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.
Jin
Liang
L
Institute of Neuroscience, Fourth Military Medical University, Xi'an 710032, China. jin173@hotmail.com
Jin
Wei-lin
WL
Ju
Gong
G
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005260:Female; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D011247:Pregnancy; D018045:Receptors, Oxytocin / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
D018045:Receptors, Oxytocin; D011994:Recombinant Proteins