人突变型p53及EB病毒LMP1转基因小鼠建立.docVIP

人突变型p53及EB病毒LMP1转基因小鼠建立 　 作者：何迎春，田道法，卢芳国，毛积芳 【关键词】 鼻咽癌前病变 Construction of transgenic mice containing human mutant p53 and EB virus latent membrane protein 1 　　【Abstract】 AIM: To establish bitransgenic mice models containing mutation p53 gene and EpsteinBarr (EB) virus latent membrane protein 1 gene （LMP1） and to provide reliable tools for studying the reciprocity of mutant p53 with LMP1 in the tumorogenesis process of nasopharyngeal precancerosis. METHODS: Molecular cloning techniques were used to construct the recombinant plasmid pLMP1p53mt containing mutant p53 gene and EB virus LMP1 gene. Linear recombinant plasmid pLMP1p53mt was transfected into mouse androgenesis of germ cells by microinjection and then survival germ cells treated were planted into fallopian tubes of artificial pregnant female mice. Threeweek filial generation mice were screened by PCR and further identified by southern blot, and the histopathological characteristics of different tissues in 4month transgenic mice were observed by HE staining. RESULTS: Seven hundred and seven mouse zygotes were treated with transgenic vectors DNA by microinjection and then transplanted into 30 artificial pregnant female mice. Twentysix of these mice were gravid. The transplantation rate was 86.67%. A total of 179 F0 mice were obtained, of which 9 were carried p53mt gene and LMP1 gene screened by PCR. The positive rate of transgenic mice was 5.03% (9/179). The 9 positive F0 mice carrying foreign genes were further confirmed by southern blot. The rhinal mucosa, nasopharyngeal tissues and oesophagus of one founder had inflammation and its nasopharynx mucosa had focal hyperplasia. CONCLUSION: Transgenic mouse models integrating human mutant p53 gene and LMP1 and with focal hyperplasia of nasopharynx mucosa have been established by microinjection, which provides an effective model in studying the mechanism of nasopharyngeal precancerous lesions. 　　【Keywords】 mutation p53 gene; latent membrane protein 1 gene; nasopha