Development of a yeast-based recombination cloning/system for the analysis of gene products from diverse human immunodeficiency virus type 1 isolates.
A recent shift from studies on a few subtype B laboratory human immunodeficiency virus type 1 (HIV-1) clones to analyses of extremely diverse primary HIV-1 isolates from different subtype requires the development of a rapid and generic cloning technique. This report describes the use of gap repair/recombination in yeast to shuttle env, gag, and pol genes from diverse HIV-1 subtypes into a DNA vector that can be amplified in bacteria and can express the gene of interest in mammalian cells. These diverse HIV-1 genes have also been introduced into an infectious clone to produce chimeric viruses that are useful for studies on drug susceptibility, receptor binding and fitness.
Marozsan
Andre J
AJ
Division of Infectious Diseases, Department of Medicine, Case Western Reserve University, BRB 1029, 10900 Euclid Avenue, Cleveland, OH 44106, USA.
Arts
Eric J
EJ
D002460:Cell Line; D003001:Cloning, Molecular; D015686:Gene Products, env / Q000235:genetics / Q000378:metabolism*; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism*; D015684:Gene Products, pol / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D010957:Plasmids; D011995:Recombination, Genetic; D015003:Yeasts / Q000235:genetics
D015686:Gene Products, env; D015683:Gene Products, gag; D015684:Gene Products, pol