Human Immunodeficiency Virus 유전자 조작 및 양서류 난자에서의 발현에 관한 연구 = The Expression of Subcloned Human Immunodeficiency Virus Genes Microinjected into the Amphibian Oocyte

다국어 초록 (Multilingual Abstract)

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.