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ＥＢ病毒（ＥＢＶ）是一种与地区性伯基特氏淋巴瘤、鼻咽癌、何杰金氏病等多种人体肿瘤有关的疱疹病毒．已往的研究表明，潜伏膜蛋白（ＬＭＰ）基因是ＥＢＶ最可能的致瘤基因．为制备ＬＭＰ基因转基因小鼠，探讨ＬＭＰ的体内致瘤作用，首先构建了含鼠金属硫蛋白－１（ＭＴ－１）基因调控区和ＬＭＰ基因编码区的ｐＢＲ３２２－ＭＴ－ＬＭＰ质粒，并用电击法将该质粒与ｐＫＪ１－Ｎｅｏ质粒共转染人胃癌细胞株ＭＧＣ，对ＭＴ－ＬＭＰ基因在转染细胞中的整合、转录情况及重金属镉和镍对该融合基因的转录调控进行了研究．结果表明：（１）两质粒共转染效率为８６．７％；（２）ＰＣＲ和Ｓｏｕｔｈｅｒｎ杂交分析显示，完整的ＭＴ－ＬＭＰ基因已整合入转染的ＭＧＣ细胞基因组，且在不同的转染细胞克隆中，ＭＴ－ＬＭＰ基因整合的方式及拷贝数不同，拷贝数从１到１９不等；（３）ＲＴ－ＰＣＲ和Ｎｏｒｔｈｅｒｎ杂交分析证实，ＭＴ－ＬＭＰ基因不仅在转染的ＭＧＣ中能够转录，而且在１０μｍｏｌ／Ｌ镉诱导下，ＭＴ－ＬＭＰ基因转录增强，平均增高约１．４倍．结果说明，在ＭＴ－１基因调控区指导下，ＬＭＰ基因不但有ｍＲＮＡ水平的表达，而且其表达受重金属镉的调控，上述结果为制备ＭＴ－ＬＭＰ转基因小鼠?
Epstein-Barr virus (EBV) is a herpesvirus that is associated with a variety of human tumors, including Burkitt’s lymphoma, nasopharyngeal cancer, Hodgkin’s disease and others. Previous studies have shown that latent membrane protein (LMP) gene is EBV most likely oncogene. To prepare LMP gene transgenic mice and investigate the in vivo tumorigenicity of LMP, pBR322-MT-LMP plasmids containing murine metallothionein-1 (MT-1) gene regulatory region and LMP gene coding region were constructed and electroporated The plasmid and pKJ1-Neo plasmid co-transfected human gastric cancer cell line MGC, MT-LMP gene transfected cells in the integration and transcription of heavy metals and cadmium and nickel on the transcriptional regulation of the fusion gene was studied. The results showed that: (1) the efficiency of co-transfection of two plasmids was 86.7%; (2) PCR and Southern hybridization analysis showed that the complete MT-LMP gene was integrated into the genome of transfected MGC cells, In the cell cloning, the way of MT-LMP gene integration and copy number is different, and the copy number varies from 1 to 19. (3) RT-PCR and Northern blot analysis confirmed that MT-LMP gene not only transcribed in transfected MGC , And the transcription of MT-LMP gene was enhanced with an average increase of 1.4-fold under the induction of 10μmol / L cadmium. The results showed that, under the guidance of MT-1 gene regulatory region, not only LMP gene mRNA expression, and its expression by heavy metal cadmium regulation, the above results for the preparation of MT-LMP transgenic mice?