Author(s): YAO Qi, WANG Ai-yan, LING Min, Department of Biochemistry and Molecular Biology, Guangxi Medical University
Pages: 60-62
Year: 2014 Issue:  4
Journal: Biotechnology
Keyword:  Mycobacterium avium; Identification; Genomic library;
Abstract: Objective: To construct genomic library of Mycobacterium avium. Method: 15 cases of sputum Mycobacteria cultures positive specimens in HIV /AIDS patients were identified by 16 S rRNA gene sequencing. Total DNA extracted from M. avium was digested by Sau3AⅠ,and then was ligated to pUC19 vector digested with Bam HⅠ to construct genomic library when the ligated product was transformed into E. coli JM109. The quality of the genomic library was evaluated by titer calculation and clones restriction analysis. Result: In 15 cases of Mycobacteria,9 cases of M. tuberculosis,4 cases of M. avium and 2 cases of M. abscessus were found. The titer of the genomic library was estimated to be 2. 58 × 105 cfu /ml. Insert fragments( 0. 5- 1 kb) could be seen when 9 clones selected randomly from the genomic library were digested with BamHⅠ. Conclusion: A genomic library of M. avium was construted,it provides an effective tool to investigate the structures and function of pathogenic genes in M. avium.