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  本文關鍵詞：銅綠假單胞菌噬菌體D204的生物學特性和基因組學研究　出處：《上海交通大學學報(醫(yī)學版)》2016年01期 　論文類型：期刊論文

  更多相關文章： 銅綠假單胞菌 噬菌體 基因組 生物學特性

【摘要】：目的針對銅綠假單胞菌多耐藥菌的不斷出現(xiàn),尋找高效、廣譜的銅綠假單胞菌噬菌體。方法以醫(yī)院臨床上分離到的銅綠假單胞菌為宿主菌,從環(huán)境污水中分離純化銅綠假單胞菌噬菌體,磷鎢酸負染法電鏡觀察噬菌體的形態(tài)和大小,測定其效價、宿主譜、酸堿穩(wěn)定性和熱穩(wěn)定性,繪制一步生長曲線、吸附曲線和感染曲線,并抽取噬菌體DNA進行測序及分析。結果分離到1株銅綠假單胞菌噬菌體,并命名為D204。D204在pH值為4~11及溫度37~50℃環(huán)境下具有較高活性,電子顯微鏡觀察發(fā)現(xiàn)此噬菌體由直徑約為50 nm的多面體頭部和約15 nm的短尾組成。一步生長曲線表明噬菌體在裂解宿主菌時,潛伏期為30 min,爆發(fā)期為40 min,裂解量約為160 pfu/cell。吸附曲線顯示50 min時,其吸附率達95%以上。感染曲線顯示當感染復數(shù)為0.1和1時,對宿主菌生長有抑制作用,在180 min后可完全抑制宿主菌生長。其基因組為環(huán)狀雙鏈DNA,全長為50 609 bp,GC含量為44.8%。預測共有75個開放閱讀框。結論分離、篩選出的銅綠假單胞菌噬菌體D204具有較強的裂解宿主的能力,其對酸堿和熱比較穩(wěn)定,裂解宿主時潛伏期較短,可以作為銅綠假單胞菌的儲備噬菌體。
[Abstract]:Objective to search for efficient and broad-spectrum Pseudomonas aeruginosa bacteriophage. Methods Pseudomonas aeruginosa isolated clinically in hospital was used as host bacteria. Pseudomonas aeruginosa phage was isolated and purified from environmental sewage. The morphology and size of phage were observed by phosphotungstic acid negative staining electron microscope. The titer, host spectrum, stability and thermal stability of the bacteriophage were determined, and the one-step growth curve was drawn. The adsorption curve and infection curve were obtained. The phage DNA was sequenced and analyzed. The results showed that a bacteriophage strain of Pseudomonas aeruginosa was isolated. It was named D204.D204 with high activity at pH 4? 11 and temperature 37? 50? 鈩，

本文編號：1413718