Cloning of HBx,Ras Gene and Preliminary Study on Establishing Transgenic Mice by Transfection of Spermatogonial Cells

Objective:A study was performed to investigate the possibility of using transfection of spermatogonial cells to estab- lish transgenic mice with hepatitisB virus X gene and mutated ras gene.Methods:The eukaryotic expression vectors of pcDNA3.1-HBx and pEGFP-N1-Kras were constructed.The mixture of two recombinant vectors coated with liposome was injected into the testicle tissue of 4 male mice.The treated male mice were fertilized with females four weeks later.The tails of 2～4 weeks old first generation mice were cut and polymerase chain reaction was performed to detect integration of the target gene into the genomic DNA of the mice.Results:35 first generation mice were born and 3 were PCR positive(positive rate is 8.6%).Among the 3 ones,2 were positive for HBx,1 was posi- tive for both HBx and ras.Conclusion:It is practical that using the spermatogonial cells as the vector to produce the transgenic mice of HBx and ras gene,and could play a role in establishing transgenic animal model of tree shrews.