HIV의 gag-pol 유전자를 가진 plasmid의 제작이 가능하였으며 이들 plasmid들은 감염의 위험성이 없고 유전자의 발현은 가능하였다. 또한 HIV 자체의 LTR보다 SV40 promoter가 유전자의 발현에 더 강력한 것을 알았으며 양서류 난자에서는 transactivator인 tat 유전자 없이도 HIV 유전자의 발현이 되었다. Xenopus 난자에서 HIV의 gag-pol 유전자들의 발현이 가능하였으므로 이들 유전자들을 이용하여 transgenic mouse의 제작이 가능하다고 생각된다. 이러한 transgenic mouse는 HIV연구에 좋은 실험 동물 model이 될 것이다. 한편 양서류 난자에 미세주입된 DNA에 의한 RNA와 단백질 합성 시스템은 외부유전자의 발현 여부를 검정하는데 유용한 도구가 될 것이다.

This study was undertaken to elucidate the expression of recombinant gag-pol gene of human immunodeficiency virus(HIV), the virus which causes aquired immune deficiency syndrome(AIDS), in the Xenopus oocytes. pHXB-2D containing complete HIV provirus was digested with restriction endonucleases, Sal Ⅰ and Xho Ⅰ. The cleaved DNA fragments containing gag-pol gene and long terminal repeat sequence(LTR) were ligated to construct recombinant pHXGP plasmid. pHXGP was treated with restriction endonuclease, Sac Ⅰ. The cleaved DNA fragment containing gag-pol gene was recombined with pSVL vector containing SV40 promoter to construct pSSGP plasmid. After microinjection of pHXGP and pSSGP plasmids into Xenopus oocytes, 32P-GTP was injected into the oocyte cytoplasm to assess RNA synthesis. The RNA synthesis by pHXGP and pSSGP plasmids was confirmed respectively and the RNA synthesis by pSSGP plasmid containing SV40 promotor increased more than that by pHXGP plasmid. The protein synthesis by both plasmids was assessed after microinjection of 35S-methionine into Xenopus oocytes and was confirmed respectively by immunoelectrophoresis using monoclonal antibodies of p24 and reverse transcriptase. The protein synthesis by pSSGP also increased more than that by pHXGP plasmid. The results showed that the construction of plasmid containing gag-pol gene of HIV was possible and the plasmids were noninfectious, but the genes were expressed. The expression of genes by SV40 promoter was more active than that by LTR of HIV. The expression of HIV genes without tat gene was also confirmed in amphibian oocyte.