关键词: 比格犬, 慢病毒载体, 生殖细胞

Abstract: Abstract: Objective To observe the transfection efficiency of lentiviral vector for dog testicular germ cells so as to provide the foundation for generating the transgenic animal by lentiviral vector-based sperm-mediated gene transfer． Method LV-GFP with the titers 5 × 108 ，1 × 108 ，2 × 107 TU / mL was injected respectively into the bilateral testis of Beagle dogs in high，medium and low-dose group at a dose of 2 ml / dog． Sperm-collecting was set about from the second week after the testicular injection． The integration and expression of GFP gene were assessed by PCR tests of semen DNA and sperm detection with fluorescence microscope． Results 1) In high-dose group the integration and expression of GFP gene firstly appeared at the 4th week and maintained till the 17th week，and it appeared at the 5th week in the medium and low-dose group and lasted till to the 14th and 12th week respectively; 2) The peak level of GFP expression occurred in 7 to 10 weeks after the testicular injection; 3) GFP expressed in the entire sperm，but expressed highly in the acrosomal back zone and the neck section of sperm tail; 4) There was ascent trend of dog sperm deformity rate in high and low-dose group during 2 weeks to 6 weeks after the injection; 5) During the peak period of GFP expression，the percentage of GFP sperm in high，medium and low-dose group was respectively 43. 33% ，35. 53% ，4. 55% ，with a significant difference． Conclusion The lentiviral vectorbased SMGT can successfully generate transgenic dog sperm，and the transgenic rate，the intensity and the duration of transgenic expression were related to the titer of lentiviral vector．

Key words: Beagle dog, lentiviral vector, germ cell