DSPP G0 generation of transgenic mice to obtain.docVIP

 PAGE \* MERGEFORMAT 9 DSPP G0 generation of transgenic mice to obtain Author: Sun Hantang, Xiao Zhen, Wu replacement, Xu Jiang, Jian Fei [Keywords:] DSPP Generation of transgenic mouse founders of DSPP [Abstract] AIM: To establish tetracyclineresponsive regulated transgenic mice of dentin sialophosphoprotein (DSPP). METHODS: DSPP coding sequence was contained in plasmid pBluescriptDSPP and the transgenic vector was constructed by subcloning it into pTRE2 by NotI / SalI digestion. Following identification by enzyme digestion and sequencing, the final plasmid was named pTRE2DSPP. After linearized by XhoI and recovery, the plasmid was microinjected into the male pronucleus of the zygotes and the zygotes in good condition were implanted into pseudopregnant mice. The tail DNA of 3 weekpups was tested by PCR and Southern blot. RESULTS: Six hundred and fiftyseven embryos were implanted to 28 recipient pseudopregnant mice, with 6 of the 85 pups carrying the transgene. The establishment of the transgenic line progressed from the 6 positive ones. CONCLUSION: The founders of the tetracyclineresponsive regulated transgenic mice of DSPP are established successfully. [Keywords] tooth; dentin sialophosphoprotein; mice, transgenic; microinjections [Abstract] Objective: To construct tetracycline-controlled nature of DSPP (DSPP) transgenic mice with the Department of the controlled system. Methods: From the plasmid pBluescriptDSPP in the mouse DSPP coding sequence was subcloned into pTRE2 the NotI / SalI sites , the construction of genetically modified vector by restriction enzyme digestion and sequencing confirmed, named pTRE2DSPP; the XhoI digested linearized vector DNA was purified, micro-injected into mouse eggs of the male pronucleus, the selection of the growth state of good fertilization eggs transplanted into pseudo-pregnant female mouse oviduct. earners mice 3 wk after birth, using PCR and Southern Blot test positive in mice. Results: A total of survival of ferti