大肠杆菌肽基脯氨酰顺反异构酶slyD基因缺失株的构建及生物学特性研究  

Construction and biological characteristics of peptidyl-prolyl cis-trans isomerase sly D gene defective mutant of Escherichia coli

作　　者：周薇[1,2] 刘松艳[2] 王曼宇 衣菲[1,2] 田秋丰[1,2] 陈志宝[1] 刘思国[2] 于申业[2] 

机构地区：[1]黑龙江八一农垦大学生命科学技术学院,黑龙江大庆163319 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/动物细菌病研究室,黑龙江哈尔滨150001 [3]吉林农业大学动物科学技术学院,吉林长春130118

出　　处：《中国预防兽医学报》2016年第1期6-10,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金项目(31470893;31272538);863计划(2011AA10A210);国家重点基础研究发展计划[973计划(2012CB518801)];国家公益性行业(农业)科研专项(201403054)

摘　　要：肽基脯氨酰顺反异构酶(PPIases)广泛存在于原核生物中,主要生物功能是专一性催化已磷酸化的丝/苏-脯氨酞顺/反异构。为研究大肠杆菌PPIases sly D基因缺失对其生物学特性的影响,本研究利用改进的Red重组方法构建大肠杆菌BL21株的sly D缺失株,命名为BL21Δsly D;并通过突变株的形态学变化、生长特性、生化代谢特性、耐酸性、耐渗透压能力、耐温度缺氧能力对其进行鉴定。结果表明,缺失株生长速率、耐高温缺氧能力与亲本株相比差异不显著,然而突变株的耐酸性与耐渗透压能力则显著低于亲本株(p<0.01),表明大肠杆菌PPIases sly D基因可能影响双组份调控系统的调节作用。本研究构建的大肠杆菌sly D基因缺失株BL21Δsly D为进一步研究大肠杆菌sly D基因的功能奠定了基础。The peptidyl-prolyl cis-trans isomerase （PPIases） is commonly found in prokaryotes, which specifically isomerizes the phosphorylated Ser/Thr-Por bonds in certain proteins. In order to investigate the biological characteristics of the PPIases subunit encoding slyD gene defective strain of Escherichia coli, the slyD gene was knocked out from the chromosome of BL21 （DE3） pLysS strain using an improved Red homologous recombination system, and the mutant was named BL21AslyD, which was identified by observing the morphological changes of slyD-deleted mutant, analyzing its growth characteristics, biochemical metabolism characteristics, acid stress, osmotic stress and temperature stress. The results showed that the growth rate and the temperature stress of the BL21As/yD had no significant difference to the parental strain; but the acid stress and osmotic stress was significant lower than parental strain, suggesting that the PPIases smight affect the function of two-component system in E.coli. In conclusion, the constructed slyD defective mutant of E.coli provided a basis for further study on the function of PPIases in E.coli.

关 键 词：大肠杆菌 肽基脯氨酰顺反异构酶 生物学特性 环境压力 

分 类 号：S852.61[农业科学—基础兽医学]