Generation of transgenic mouse founders of DSPP

    SUN Hanª²Tang, XIAO Mingª²Zhen, WU Buª²Ling, X¦a Guoª²Jiang, FEI Jian

    1Department of Oral Medicine, College of Stomatology, Fourth Military Medical University, Xi¬ðan 710033, China, 2Shanghai Nanfang Research Center of Biomodel Organism, Shanghai 201203, China, 3Shanghai Institute of Biochemistry and Cell Biology, Science Academy of China, Shanghai 200031, China

    ¡¾Abstract¡¿ AIM: To establish tetracyclineª²responsive regulated transgenic mice of dentin sialophosphoprotein (DSPP). METHODS: DSPP coding sequence was contained in plasmid pBluescriptª²DSPP 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 pTRE2ª²DSPP. 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 weekª²pups was tested by PCR and Southern blot. RESULTS: Six hundred and fiftyª²seven 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 tetracyclineª²responsive regulated transgenic mice of DSPP are established successfully. ......