Studies on renin-2 gene in transgenic rats

Objective To determine the function of, in vivo, renin and its role in the pathogenesis of hypertension. Methods A renin-2 gene restriction map was constructed by endonuclease dige stion and Southern blotting hybridization. Transgenic rats were produced vi a microinjection method. Results The 24 kb fragments containing mouse full-length ren-2 and its flan king sequence were cleaved by single enzymes (EcoR I, Kpn I and BamH I) and combined enzymes (EcoR I/Kpn I, Kpn I/BamH I and BamH I/EcoR I), respectiv ely. The digests were electrophoresed in 0.8% agarose plates and transferre d onto NC membranes. Radioactive 735 bp and 1400 bp probes obtained from ha lf and full-length renin-l cDNA were used in southern blotting hybridizatio n. According to the electrophoresis and hybridization patterns, a ren-2 res triction map was constructed. 1603 fertilized rat ova after injection with purified 24 kb renin-2 gene were implanted into the oviducts of 81 pseudopr egnant recipients in about 20 ova per female rat. 306 progenies were obtain ed from 50 foster mothers (average of pregnancies was 56.6%). 248 survived pups were identified by PCR analysis and Southern hybridization, and eight positive rats were found to be the transgenic rats (founder, Fl. All of the m carried long fragments (24 kb) of renin-2 gene with normal blood pressure . Preliminary breeding and screening were carried out in the founder. Total survival pups (17.8%) and overall efficiencies (1%) were harvested as the same as those reported in the literatures. A systemic observation and the p roblems occurred during production of transgenic rats were also described b esides the technique procedure used in this study. Conclusions Mapping of full-length murine ren-2 can be used in investigatio n of the structure and function of the gene. The results denoted that the r en-2 transgenic rats were successfully established in this study and the te chnique used in the production of transgenic rats was proved to be valid in leading to wide-spread application of transgenic technique to many other r elated researches.