论文部分内容阅读
目的 :分析乙型肝炎病毒 pre S2蛋白在 3′末端缺失的 pre S/ S基因转基因小鼠肝脏中的分布及其病理学作用。方法 :采用原核显微注射法将质粒 pc DNA3.1- pre S/ St注射入小鼠受精卵雄原核 ,制备转基因小鼠。PCR法在基因组水平筛选 3′末端缺失的 pre S/ S基因转基因小鼠首建者 (founder)及后代 ;免疫组织化学法在蛋白水平检测 pre S2蛋白在这些小鼠中的表达 ;H- E染色分析转基因小鼠肝组织的病理学变化。结果 :经原核显微注射法将目的片段注射入受精卵雄原核后 ,共出生 15只新生小鼠 ,其中存活 7只 ,经 PCR检测后获得 2只 founder转基因小鼠 ,命名为 C5 7- Tg N (pre S/ St) SMMU。免疫组织化学检测发现转基因小鼠肝细胞质中有 pre S 2蛋白表达 ,H- E染色发现转基因小鼠肝组织中央静脉周围有淋巴细胞聚集。将这 2只转基因小鼠与正常同系异性小鼠交配 ,进行传代培育 ,PCR法筛选阳性转基因小鼠 ,目前已传至 F2 代。 结论 :本研究成功建立了稳定遗传 3′末端缺失的 pre S/ S基因并表达 pre S2蛋白的转基因小鼠 C5 7- Tg N((pre S/ St) SMMU,它将是体内研究 3′末端缺失的 pre S/ S基因的表达产物在体内的生物学作用及其与肝细胞内细胞癌基因的转录激活之间关系的理想动物模型
OBJECTIVE: To analyze the distribution and pathological role of hepatitis B virus pre-S2 protein in the liver of 3’-terminal pre S / S transgenic mice. Methods: The plasmid pcDNA3.1-pre S / St was injected into the pronucleus of mouse fertilized eggs by pronuclear microinjection to prepare transgenic mice. PCR was used to screen the founder and offspring of pre S / S gene transgenic mice lacking the 3 ’end at the genome level. The expression of pre-S2 protein in these mice was detected by immunohistochemistry at the protein level. H-E Staining Analysis of Pathological Changes in Liver of Transgenic Mice. Results: Fifteen newborn mice were born after fertilized eggs were injected into prokaryotic nucleus by pronuclear microinjection. Seven of them were viable, and two transgenic mice were obtained after PCR. N (pre S / St) SMMU. Immunohistochemistry showed that there was preS 2 protein expression in the liver cytoplasm of transgenic mice. H-E staining showed lymphocyte aggregation around the central vein of the transgenic mice. The two transgenic mice were crossed with normal male mice and subcultured. The positive transgenic mice were screened by PCR, which has been reported to F2 generation. CONCLUSIONS: The present study successfully established a transgenic mouse C5 7-Tg N ((pre S / St) SMMU that stably inherits the 3 ’end deleted pre S / S gene and pre S2 protein and will be an in vivo study of the 3’ end The biological role of the expression products of the deleted pre S / S gene in vivo and its ideal animal model for the relationship with the transcriptional activation of intracellular hepatocellular carcinoma genes