Wu Jin ming, Lin Ju-sheng, Chen Bi-tao, Zheng Xin-min, Zhao Hao-bin, Liang Kuo-huan
Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong Science and Technology University, Wuhan 430030, China.
Zhonghua Gan Zang Bing Za Zhi. 2003 Jun;11(6):338-40.
To establish a highly expressing and replicating hepatitis B virus (HBV) genome transgenic mouse models for screening anti-HBV drugs and investigating the pathogenesis of hepatitis B.
Elongated HBV genome as the investigated gene was transducted into the pronuclei of the fertilized eggs of mice by the technique of microinjection, then the eggs were transplanted into the oviducts of the pseudopregnant mice. All the newborn mice were screened and identified by PCR and Southern blot detecting genomic DNA in tail tissue, then the positive mice were examined plasma HBsAg, HBeAg by ELISA and plasma HBV DNA by Southern blot.
Among the 61 offsprings, 18 were positive for tail tissue HBV DNA examination, 7 of which were positive for replication and expression detection.
Transgenic mice with elongated HBV genome possess high efficiency of replication and expression, which can be used for further investigation.
建立高表达和复制的乙型肝炎病毒(HBV)基因组转基因小鼠模型,用于筛选抗HBV药物及研究乙型肝炎发病机制。
采用显微注射技术将延长的HBV基因组作为研究基因导入小鼠受精卵原核,然后将受精卵移植到假孕小鼠输卵管。所有新生小鼠通过PCR和Southern印迹检测尾组织基因组DNA进行筛选和鉴定,然后用ELISA检测阳性小鼠血浆HBsAg、HBeAg,用Southern印迹检测血浆HBV DNA。
61只子代中,18只尾组织HBV DNA检测为阳性,其中7只复制和表达检测为阳性。
延长HBV基因组转基因小鼠具有较高的复制和表达效率,可用于进一步研究。