[1]孙卫忠,胡晓梅,饶贤才,等.铜绿假单胞菌噬菌体PaP1内溶素基因克隆、表达及活性分析[J].第三军医大学学报,2008,30(13):1268-1271.
 SUN Wei-zhong,HU Xiao-mei,RAO Xian-cai,et al.Cloning, expression and enzymatic assay of bacteriophage PaP1 endolysin against Pseudomonas aeruginosa[J].J Third Mil Med Univ,2008,30(13):1268-1271.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第13期
页码:
1268-1271
栏目:
论著
出版日期:
2008-07-15

文章信息/Info

Title:
Cloning, expression and enzymatic assay of bacteriophage PaP1 endolysin against Pseudomonas aeruginosa
作者:
孙卫忠胡晓梅饶贤才谭银玲陈志谨丛延广胡福泉
第三军医大学基础医学部微生物学教研室,重庆市微生物工程实验室
Author(s):
SUN Wei-zhong HU Xiao-mei RAO Xian-cai TAN Yin-ling CHEN Zhi-jin CONG Yan-guang HU Fu-quan
Department of Microbiology, College of Medicine, Third Military Medical University, Chongqing 400038, China
关键词:
噬菌体内溶素基因表达活性鉴定
Keywords:
bacteriophage endolysin gene expression enzymatic activity
分类号:
Q939.48; R378.991; R394-33
文献标志码:
A
摘要:
目的  构建铜绿假单胞菌噬菌体内溶素重组原核表达载体,检测重组蛋白的酶学活性及抑菌活性。  方法  利用重组技术将铜绿假单胞菌裂解性噬菌体PaP1内溶素基因构建到表达载体pQE31中,并在大肠杆菌M15(pREP4)中诱导表达,通过非变性方法纯化重组融合蛋白,利用酶谱电泳、抑菌实验等技术检测酶学活性及抑菌活性。  结果  酶谱电泳方法结果显示重组融合蛋白对铜绿假单胞菌细胞壁肽聚糖有降解作用,抑菌活性检测结果显示重组融合蛋白对金黄色葡萄球菌有一定的抑菌效果。  结论  该研究从实验方面证实了噬菌体PaP1内溶素基因的生物学功能。
Abstract:
Objective    To construct, express and purify recombinant endolysin of Pseudomonas aeruginosa (P. aeruginosa) bacteriophage PaP1, and detect its enzymatic activity and antibacterial activity.     Methods    The objective endolysin fragment of bacteriophage PaP1 was amplified from the genome, and then cloned into the expression vector pQE31. Then the fusion protein expressed in E.coli M15(pREP4) and purified under native conditions. The enzymatic activity was detected by zymography assay and its biological activity was detected by antibacterial activity assay.     Results    Enzymatic assay showed that the recombinant protein could cause the lysis of the purified P. aeruginosa peptidoglycan. Antibacterial activity assay showed that the recombinant protein inhibited the growth of Staphylococcus aureus.     Conclusion    The obtained phage endolysin exerts antibacterial activity to P. aeruginosa.

参考文献/References:

孙卫忠,胡晓梅,饶贤才,等. 铜绿假单胞菌噬菌体PaP1内溶素基因克隆、表达及活性分析[J]. 第三军医大学学报, 2008, 30(13):1268-1271.

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更新日期/Last Update: 2008-06-30