用精原细胞介导法制备乙肝病毒X基因转基因小鼠的实验研究

目的：探讨精原细胞介导法制备乙肝病毒X基因转基因小鼠的可行性。方法：构建乙肝病毒X基因表达载体pcDNA3-X;应用微量注射针将脂质体包裹的pcDNA3-X表达载体直接注入C57BL／6N雄性小鼠睾丸的曲细精管内，于第一次注射后6周，与雌鼠交配，用PCR法和免疫组化法检测仔鼠基因组中的外源DNA和肝组织中表达的pX蛋白。结果：测序结果与文献报道一致，并且X基因在Hela细胞中表达；共产仔鼠16只，其中1只为阳性仔鼠，阳性率为6．25％。结论：初步证明用精原干细胞介导法制备X基因转基因小鼠是可行的。

Establishment of Transgenic Mice of Hepatitis B Virus X Gene (adr subtype) Mediated by Spermatogonial Cells

Objective To test the possibility of using spermatogonial cells as vector in the establishment of HBV X gene transgenic mice. Methods Eukaryotic expression vector pcDNA3 X of hepatitis B virus X gene was constructed. The recombinant pcDNA3 X packed by liposome was injected into testicle tissue of male C 57 BL/6N mice using mcirovolume injector. Six weeks after the first injection,the mice were crossed with female mice. Polymerase chain reaction and immunohistochemistry were applied to identify the viral DNA(HBx gene) harbored in mouse genome and the expression of HBx gene in liver tissue. Results The sequence of the X gene is consistent with that reported in literature.The X gene can be expressed in Hela cells.The positive rate in daughter mice was 6.25%. Conclusion It is possible to use spermatogonial cells as vector to establish HBx transgenic mice.