translation agency

Construction of a recombinant retroviral interfering particle containing a defective HIV-1 genome.
Federico M; Taddeo BM; Orecchia A; Saggio I; Carlini F; Verani P; Rossi
September 30, 1990
Int Conf AIDS. 1989 Jun 4-9;5:684 (abstract no. C.755). Unique

OBJECTIVE: By infecting Hut-78 cells with an RT positive supernatant of PBL from an AIDS patient, we have isolated a non-producer HIV-infected cell clone (F12), which exhibits a viral RNA pattern superimposable with that of productive cell clones, and shows resistance to HIV-1 or HIV-2 superinfection. In order to study the phenomenon of viral interference, the whole genome will be transferred in HIV-sensitive cell lines via transfection of a retroviral vector construct in an amphotropic packaging line. METHODS: From an Sst I genomic library of F12 clone we have molecularly cloned the whole provirus in pUc-19 and then subcloned it in retroviral vectors bearing the G418 resistance gene such as pLJ (MLV LTRs promoter), Ntk (thymidine kinase promoter) and NSV (SV40 promoter). RESULTS: The pLJ construct has been transfected in an amphotropic retrovirus packaging line (PA 317), in order to recover a recombinant retrovirus containing the provirus originally integrated in F12 clone. More than 30 G418-resistant PA317 clones have been obtained. Indirect IFA analysis using a pool of human positive sera was clearly positive in two PA317 clones also showing an integrated construct by Southern Blot. A more detailed molecular characterization will be discussed.

*Cloning, Molecular *Genes, Viral Genetic Vectors HIV-1/*GENETICS Retroviridae/*GENETICS *Transfection ABSTRACT