摘要
目的:利用显微注射方法产生整合庚型肝炎病毒(HGV)全基因的转基因小鼠,并研究HGV基因的遗传特性。方法:将含有HGV全长基因的载体线性化后,将目的基因DNA溶于显微注射用缓冲液,依常规方法进行显微注射,得到仔鼠后用PCR及基因组DNA Southern印迹法鉴定。阳性小鼠即为建立者(founder)小鼠,将founder小鼠与正常小鼠交配得到F代小鼠,随后将F1代阳性鼠与正常小鼠交配得到F2代小鼠。结果:共得到5只founder小鼠,其中4只与正常小鼠交配,得到F1代小鼠共41只,其中29只为阳性,阳性率为71％。F2代小鼠共21只,其中16只为阳性,阳性率为76％。结论:得到了整合有HGV全长基因的转基因小鼠,并证明HGV基因可以在转基因小鼠体内稳定传递。
Objective: To establish a transgenic mouse model carrying full-length hepatitis G virus (HGV) genome for HGV related studies. Methods: Plasmid pHGVqzl8-10 containing full-length HGV genome was linearized with Not I and Xba I ,then the target DNA fragment was microinjected into the pronuclei of fertilized eggs of donor mouse with the conventional methods. Tail tissue DNA from the transgenic mouse was amplified by PCR with oligonucleotide primers from HGV 5'NCR region. To further characterize the transgenic founders,Southern blot was performed with probe of HGV NS5 region,and then the founders and positive F1 mouse were mated with normal mice of the same line. Results: Five transgenic founders were obtained,and 4 of them were mated with normal mice of the same line. The positive ratio was 71% in F1 generation,76% in F2 generation. Conclusion: Transgenic mice harboring full-lengh HGV genome are obtained from present study. The exogenous HGV gene can be transmitted through transgenic germ line.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2002年第11期1175-1178,共4页
Academic Journal of Second Military Medical University
基金
This work is supported by the National Natural Scienee Foundation of China(No.39970394,39825116).
