奶牛瘤胃微生物BAC文库中脲酶克隆的筛选与分析  

Screening and analysis of urease clones from rumen bacterial artificial chromosome(BAC) library

作　　者：赵圣国[1,2] 王加启[1] 卜登攀[1] 刘开朗[1] 李旦[1] 于萍[1] 魏宏阳[1] 周凌云[1] 李发弟[2] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所／动物营养学国家重点实验室,北京100193 [2]甘肃农业大学动物科学技术学院,兰州730070

出　　处：《中国农业大学学报》2008年第6期61-65,共5页Journal of China Agricultural University

基　　金：国家科技支撑计划(2006BAD12B08);国际科技合作重点项目(2006DFB32160)

摘　　要：利用脲酶选择性培养基,从奶牛瘤胃微生物BAC文库的15 360个克隆中筛选得到了12个脲酶克隆。利用酚-次氯酸法对脲酶克隆子的酶学性质分析,表明均具有不同的尿素分解能力,酶的活力范围是30～2 466 U/mg。脲酶阳性克隆插入片段大约是60 kb,限制性内切酶HindⅢ酶切图谱表明,各个克隆具有不同DNA片段,其中脲酶Urease 1、Urease 3、Urease 4和Urease 9的最适pH8.0,最适温度为60℃。本研究从奶牛瘤胃微生物BAC文库中筛选到了具有脲酶酶活特性的克隆。Using urease segregation agar, twelve urease positive clones were obtained from the rumen bacterial artificial chromosome library containing 15 360 clones. They had different urea hydrolysis activities, ranging from 30 to 2 466 U/mg based on phenol-hypochlorite urease assay. The insert size in urease positive clones was about 60 kb. The electropherogram of Hind 111 digested urease positive clones revealed that they had different DNA fragments. And the optimum pH and temperature of Urease 1, Urease 3, Urease 4 and Urease 9 clones were 8.0 and 60 ℃ respectively.

关 键 词：瘤胃微生物 细菌人工染色体文库 筛选 脲酶 酶学性质 

分 类 号：Q55[生物学—生物化学] Q785