EB病毒早期基因BHRF1转基因小鼠纯合子建立及建系

GENERATION OF HOMOZYGOTE TRANSGENIC MICE OF EPSTEIN-BARR VIRUS EARLY GENE BHRF1

目的构建整合有EB病毒(EBV)早期基因BamHI-H右向读码框1(BHRF1)基因纯合子转基因小鼠。方法通过显微注射技术构建BHRF1阳性首建鼠F0共16只,将转基因小鼠与正常小鼠交配,得到子代F1小鼠,PCR技术检测子鼠鼠尾组织DNA,鉴定筛选BHRF1阳性鼠。阳性鼠再与同系阴性鼠杂交得到F2小鼠,筛选出F2小鼠中的阳性鼠近交得F3小鼠。筛选出F3小鼠中的纯阳性鼠进行近交,得到纯阳性F4小鼠。F4小鼠再近交得到纯阳性F5小鼠。结果经PCR及测序证实,F4、F5小鼠均为BHRF1阳性鼠,测序结果经blast比对,与BHRF1基因完全相同。结论本实验首次建立了BHRF1转基因鼠模型,并成功筛选出纯阳性BHRF1转基因小鼠,为深入研究BHRF1的生物学特性及其在EBV相关肿瘤发生发展中的作用提供了理想的动物模型。

Objective To generate homozygous transgenic mice of Epstein-Barr virus early gene BHRF1. Methods Sixteen founder mice carrying BHRF1 gene were established by using microinjection. The transgenic mice mated with normal mice to generate F1. PCR were used to screen positive mice of F1. The positive F1 mice were then mated with normal mice to generate F2. The positive F2 mice were then chosen to inbreed with each other to generate F3. The pure positive mice of F3 were identified to inbreed with each other to generate F4, then F5 mice were generated by the same method. Results F4 and F5 mice were all BHRF1 positive identified by PCR and sequencing. The sequence was identical to BHRF1 gene compared by blast in Pubmed. Conclusion A BHRF1 transgenic mouse model has been successfully established and the pure mice carrying BHRF1 gene were screened for the first time, which provides an ideal animal model for further research of the biological characteristics of BHRF1 and its role in the pathogenesis of EBV-related tumors.