Resource Logo
NLM AIDSLINE

Construction of a recombinant retroviral interfering particle containing a defective HIV-1 genome.




 

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



 




Information in this article was accurate in September 30, 1990. The state of the art may have changed since the publication date. This material is designed to support, not replace, the relationship that exists between you and your doctor. Always discuss treatment options with a doctor who specializes in treating HIV.