Construction and Identification of Transgenic Mice Carrying Human Cytomegalovirus IE2 Gene

The infection of human cytomegalovirus causes several diseases of congenitally infected neonates and immunocompromised populations. The immediate–early 2 gene of human cytomegalovirus plays an essential role in viral replication and the pathogenic process. Due to the strict species-specific of HCMV infection, any animals besides human beings cannot be infected with HCMV. For this reason, few animal model which is used to explore the effect of the IE2 gene on the mRNA or protein level has been constructed to. In this study, the cDNA of IE2 mRNA (1,743 bp) was cloned into the mammalian expression vector pAV.Des1d then identified by polymerase chain reaction and sequencing analysis. Next, the expressed vector was transferred into mouse fertilized eggs from C57BL/6 mice by pronuclear microinjection to obtain the first generation of transgenic mice. In the 59 F0 generation mice, there are only 3 offspring mice were identified as IE2 positive mice. Furthermore, the positive ratios detected by PCR from F0 to F3 were 53.8%, 43.1%, 62.1%, respectively. The expression of exogenous IE2 mRNA was detected in kidney, heart, muscle, brain, spleen, and adipose tissue of F1 transgenic mice. These results suggest that we have successfully constructed a stable transgenic mouse line that can be used as a tool to explore the specific effects of IE2 gene on the growth and development of animals as well as mechanisms of its influence.