Genitic information and biological characteristics of mycobacteriophage Chy5
JIANG Li-sha1, LIU Ping3, WU Ting-ting4, GUO Shu-liang2
1. Department of Respiratory Medicine, The First Branch, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400010,China; 2. Department of Respiratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016,China; 3. Department of Respiratory Medicine, People’s Hospital of Changshou District, Chongqing 401220,China; 4. Department of Respiratory Medicine, The Third People’s Hospital of Chengdu, Chengdu 610031,China;
Abstract:To select the phage infecting Mycobacterium tuberculosis, and understand its biological characteristics and genetic information, mycobacteriophage was isolated and purified from soil. The morphological characteristics were observed by electron-microscopy, the host range was then examined by single-spot examination. Mycobacteriophage DNA was extracted. Shutgun sequences were assembled, then the gaps were filled with PCR fragments. DNAStar software Packages was used to analyze the general characteristic of genome, Glimmer 3.0 and GeneMark were applied to predict the function of the coding gene. In addition, the collinearity analysis were conducted. Lysogens was isolated to conduct an Ultraviolet induction test and super infection immunity. In results, a novel mycobacteriophage Chy5 was isolated. The head diameter was (61.5±1.4 nm) and the tail length was (114.1±2.1 nm). It could infect different clinical isolates of Mycobacterium tuberculosis, which was classified into Siphoviridae. The full length of Chy5 genome was 51 214 bp with a G+C content of 63.60%. Chy5 genome was most similar with D29, which were 88 predicted genes encoding integrase and repressor as well as attP site. The Chy5 lysogen could release free phage particles to form plaques after UV induction. Chy5 could not clear lawn of Chy5 lysogens. So Chy5 is a novel lysogenic mycobacteriophage, which could lyse mycobacterium tuberculosis and be applied to construct Luciferase reporter phages(LRP).
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