四环素调控的小鼠LAIR-1/CD305转基因小鼠的初步建立

目的：建立四环素调控的小鼠LAIR-1/CD305转基因小鼠,为进一步研究mLAIR-1分子的体内功能奠定基础.方法：构建pBI-5-mLAIR-1载体,显微注入B6D1F1受精卵,PCR检测新生小鼠基因组DNA中LAIR-1与荧光素酶（Luciferase）基因.将mLAIR-1和荧光素酶双阳性小鼠耳成纤维细胞转染含rtTA的pUHD17.1质粒,用含盐酸强力霉素（Dox）的培养基进行培养,检测细胞裂解液中荧光素酶活性.将荧光素酶表达依赖Dox小鼠与C57BL/6交配,采用PCR对子代鼠进行检测.结果：共获得9只首建鼠,其目的基因表达高度依赖Dox,并得到其中5只首建鼠的F1代小鼠.结论：获得了四环素调控的小鼠LAIR-1转基因小鼠,可用于该分子体内功能的研究.

Generation of conditional tetracycline-regulated transgenic mice overexpressing mLAIR-1

Objective:To generate the conditional tetracycline-regulated transgenic mice overexpressing mLAIR-1.Methods:The mLAIR-1 expression vector was constructed by recombination of pcDNA3-mLAIR-1 and pBI-5.Transgenic mice lines were generated by pronuclear injection by using standard techniques.Positive animals were selected based on analysis of DNA sequence from mice tail by PCR.The controlled regulation of the integrated expression unit was tested by transfecting primary mouse-ear fibroblasts obtained from DNA-positive founders with the rtTA plasmid pUHD-17.1.Founder animals that showed Dox-dependent expression of luciferase were selected and interbred with C57BL/6 mice to establish a defined genetic background.Results:We created transgenic mice in which the expression of both mLAIR-1 and luciferase cDNA were under the control of the bidirectional promoter Pbi-1.Among 13 founder animals carrying the mLAIR-1 transgene, 9 founders regulated luciferase in response to Dox as monitored in primary fibroblast cultures.5 lines were established after interbreding with C57BL/6 mice.Conclusion:The conditional tetracycline-regulated transgenic mice overexpressing mLAIR-1 have been established successfully which may provide an useful animal model for investigating the function of mLAIR-1 molecule in vivo.