Patent ID: 6261799
Filing Date: 2001-07-17
Classification: A61K,A61P,C07K,C12N

Abstract:
A method for constructing a human immunodeficiency virus type 1 (HIV-1) gp160 expression cassette, wherein said expression cassette contains a nucleic acid capable of encoding a soluble, non-cleavable, chimeric HIV-1 gp160 variant, said variant comprising the following regions:i) a first region derived from the gp160 of a first strain of HIV-1 consisting of amino acids X to Y, wherein X is a number from 1 to 271 and Y is a number from 306 to 482;ii) a second region derived from the gp160 of a second strain of HIV-1 consisting of amino acids Y+1 to the carboxyl terminus of the envelope, wherein said second region fails to contain functional major and minor proteolytic cleavage sites (amino acids 483-486 and 475-479, respectively) and functional major and minor hydrophobic domains (amino acids 487-516 and 659-680, respectively); andiii) an optional third region, present when X is greater than 1, said region derived from the gp160 of said second strain of HIV-1 and consisting of amino acids 1 to X-1,wherein the numbering scheme of X and Y is based upon the numbering scheme of the HIV-1 isolate BRU; said method comprising the following steps:i) cloning and isolating DNA fragments encoding said first, second, and third regions of the HIV-1 envelope;ii) inserting said DNA fragments into an appropriate site in an expression cassette, wherein said cassette comprises a promoter, translation initiation codon, and optional nucleic acid encoding a signal peptide in sequence and upstream of the insertion site, and a translation termination site downstream of the insertion site.