OBJECTIVE To provide effective tools for identification and characterization of M. tuberculosis genes/antigenes and to evaluate their roles in diagnosis, vaccination, drug resistance, and pathogenesis. METHODS M. tuberculosis genomic DNA obtained from H 37 Ra strain was partially digested with DNase I. The DNA fragments ranging from 4-8 kb were isolated from agarose gel and ligated to EcoR I adaptor, and the products were linked to lamda gt11 arms and packaged using an in vitro packaging extract. The different diluted bacteriophages were used to transfect bacteria Y1090. RESULTS The efficiency and titer of the library were 85% and 3 x 10(5) pfu/ml, respectively. The library contained 1.3 x 10(5) individual recombinant phage whose foreign DNA inserted fragment size was 3.5 kb on average. CONCLUSION The genomic DNA library constructed here can provide sufficient clone to cover H37Ra gene.