Induction ofMycobacterium aviumGeneExpression following Phagocytosis byHumanMacrophages

GENE EXPRESSION 477ofsuperCosl (12) fromwhich the simian virus 40 and neomy-cin resistance sequences have been removed. Portions of theamplified cDNAlibraries were cloned into XZAP11(33).Media. Mycobacterial cultures were grown in Middlebrook7H9broth,supplementedwithADCand0.05%Tween80(19).E. coli strains were grown in Lbroth (20) supplemented withthymidine, if necessary. Macrophages were cultured in RPMI1640 (17) supplementedwith2mML-glutamine and 10%freshautologous serum.Preparation of cosmid libraries. DNAwas extracted fromthe mycobacterial strains as described previously (6). Thehigh-molecular-weight chromosomal DNAwas partially di-gestedwith the restriction enzymeSau3Aandsize fractionatedas described previously (6), and fragments of 35 to 45 kbwereligated to BamHI- and XbaI-digested pYA3060 DNA. Ali-quots of the ligation mixtures were packaged in vitro by usingGigapack II Plus extracts (Stratagene Cloning Systems, LaJolla, Calif.) and transduced intoE. coli