中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 猪丹毒杆菌噬菌体裂解酶的原核表达及活性检测 畜牧与兽医 2016,48 (10) 37-40
Position: Home > Articles > Expression and activity of the lysin of Erysipelothrix rhusiopathiae bacteriophage Animal Husbandry & Veterinary Medicine 2016,48 (10) 37-40

猪丹毒杆菌噬菌体裂解酶的原核表达及活性检测

作  者：
苑文涛;张亚宁;夏雨婷;李玉峰
单  位：
南京农业大学农业部细菌学重点开放实验室
关键词：
猪丹毒杆菌;噬菌体;裂解酶;表达;活性
摘  要：
根据猪丹毒杆菌噬菌体基因测序结果,设计针对裂解酶(Ely)的特异性扩增引物,PCR扩增获得Ely产物,PCR产物经酶切后克隆入表达载体p ET-28a(+),获得重组质粒p ET-28a(+)-Ely,并转化至大肠杆菌BL21(Rosetta)中。以1mmol/L IPTG在28℃诱导8 h,SDSPAGE结果表明,蛋白在上清中高效表达;通过Ni2+-NTA亲和层析法纯化目的蛋白,其纯度在90%以上;以100、10和1μg/m L的终浓度作用于猪丹毒杆菌培养物,每隔1 h进行一次菌落计数,直至7 h,结果表明该裂解酶在体外可以有效裂解猪丹毒杆菌,并呈一定的时间和剂量依赖性。该研究将为治疗耐药性猪丹毒杆菌感染提供新的思路。
译  名：
Expression and activity of the lysin of Erysipelothrix rhusiopathiae bacteriophage
作  者：
YUAN Wentao;ZHANG Yaning;XIA Yuting;LI Yufeng;Key Laboratory of Bacteriology,Ministry of Agriculture,Nanjing Agricultural University;
关键词：
Erysipelothrix rhusiopathiae;;bacteriophage;;lysin;;expression;;activity
摘  要：
According to genome sequence of Erysipelothrix rhusiopathiae phage,PCR fragment was amplified using a pair of primers against lysin( Ely). Recombinant plasmid p ET- 28a( +)- Ely was constructed by cloning PCR fragment into p ET- 28 a and then transferred into BL21( Rosetta). After 8 hours induced by IPTG( 1 mmol / l) at 28 ℃,SDS- PAGE results showed that the target protein with the anticipated molecular weight mainly existed in cell supernatant. The purity of the protein purified by Ni2 +- NTA is more than 90%. Recombinant protein was added into E. rhusiopathiae culture at final concentration of 100,10 and 1 μg / ml,respectively. Colony forming unit( CFU)was counted from 1h to 7 h. The results indicated that Ely can effectively lyse E. rhusiopathiae in vitro in a time- and dose- dependent manner. This study will provide a new idea for treatment of drug- resistance E. rhusiopathiae.

相似文章

导出引用
计量
文章访问数: 7
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46