Author:
Thole J E,Dauwerse H G,Das P K,Groothuis D G,Schouls L M,van Embden J D
Abstract
A gene bank of Mycobacterium bovis BCG DNA in Escherichia coli was constructed by cloning Sau3A-cleaved mycobacterium DNA fragments into the lambda vector EMBL3. The expression of mycobacterial antigens was analyzed by Western blotting with hyperimmune rabbit sera. Among 770 clones tested, several were found that produced various mycobacterial antigens in low amounts, with concentrations generally close to the detection limit. One particular clone was chosen for further investigation. This clone produced a 64-kilodalton (kDa) antigen. By placing the lambda promoter PL in front of the structural gene of this antigen, an overproducing E. coli strain was obtained. Rocket-line immunoelectrophoresis experiments showed that antigens cross-reacting with the 64-kDa protein are present in a wide variety of mycobacteria and also in so-called purified protein derivatives which are routinely used for skin tests. Preliminary experiments indicate the presence of antibodies against the 64-kDa antigen in sera from tuberculosis patients.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Immunology,Microbiology,Parasitology
Reference27 articles.
1. Handbook of immunoprecipitation-in-gel techniques;Axelsen N. H.;Scan. J. Immunol. Suppl.,1983
2. Western Blotting;Burnette W. N.;Anal. Biochem.,1981
3. Relationships among mycobacteria and nocardiae based upon deoxyribonucleic acid reassociation;Bradley S. G.;J. Bacteriol.,1973
4. Screening of cloned recombinant DNA in bacteria by in sitiu colony hybridization;Cami B.;Nucleic Acids Res.,1978
5. The immunology of mycobacterial infections;Chaparas S. D.;Crit. Rev. Microbiol.,1982
Cited by
152 articles.
订阅此论文施引文献
订阅此论文施引文献，注册后可以免费订阅5篇论文的施引文献，订阅后可以查看论文全部施引文献