Objective      To provide the animal model for studying HLA-G and transplantation immunity by generating HLA-G transgenic mice with lentivirus as a vector.       Methods      A lentivirus expression vector

  FUW-HLA-G containing HLA-G encoding sequences

was constructed and 293FT cells were transfected with packaged plasmids (psPAX2 and pMD2.G) and recombinant lentivus vector plasmid FUW-HLA-G

and packed  into lentivirus. Synchronically transfected lentivus vector plasmid FUGW containing green fluorescence-marked gene was used as a control. Titer of the FUW-HLA

-G viral suspension was measured. The concentrated FUW-HLA-G viral suspension was injected into the vitellary membrane of mice murine 1-cell eggs to generate HLA-G transgenic mice. Genetic interaction in the generated HLA-G transgenic mice was identified by PCR

transcriptional expression level of transgenic gene was measured by RT-PCR and expression of HLA-G protein was detected by Western blotting.       Results      Double BamHⅠand EcoRⅠdigestion and sequencing showed that the sequences of recombinant lentivirus vector

FUW-HLA-G

were identical with its designed sequences. The concentrated and purified biological titre of FUW-HLA-G viral suspension reached over 1×10

9

TU/ml

which met the requirements of generating lentivirus transgenic mice. Six mice with positive FUW-HLA-G were screened by PCR. RT-PCR and Western blotting showed that the HLA-G was expressed in F0 and F1 transgenic mice.       Conclusion      Transgenic mice expressing HLA-G are successfully generated by lentivirus mediated gene transfer

which can thus provide an animal model for studying the function of HLA-G and its efficiency in organ or tissue transplantation.