利用双基因裂解系统制备大肠杆菌菌影的裂解效果评价  

Evaluation of the lysis effect for the preparation of Escherichia coli ghosts with the two-gene lysis system

作　　者：徐良[1] 李媛[1] 魏延宏[1] 孙颖[1] 金天明[1] 

机构地区：[1]天津农学院动物科学系,天津300384

出　　处：《黑龙江畜牧兽医》2013年第3期14-16,共3页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31072109);天津市高等学校科技发展基金项目(20060720);天津市自然科学基金重点项目(12JCZDJC22100)

摘　　要：为了比较两种裂解系统对大肠杆菌菌影的裂解差异,试验以利用双基因裂解系统制备的pBV220 DH5α、pBV-E DH5α、pBV-E-15aalinker-SN DH5α为研究对象,以pBV220 DH5α转化菌为阴性对照,并用分光光度法检测高温诱导裂解后的活细菌数。结果表明:经42℃升温诱导后菌体发生裂解,基因E-15aalinker-SN的裂解效率优于裂解基因E;在裂解过程中,Ca2+和Mg2+的添加对E蛋白的裂解作用产生干扰,诱导90 min后加入二价阳离子可以减轻干扰作用,但未见二价阳离子对于菌影形成具有促进作用。To compare the lysis differences of two types of lysis system to E. coli bacterial ghost, pBV220 DH5ct, pBV -E DH5a and pBVE -15aalinker- SN DH5a which were prepared by two- gene lysis system, were used as the research objects, taking pBV220 -transformed bacteria as a negative control, the number of living bacteria after induced lysis at high temperature was detected by spectrophotometry. The resuits showed that the bacteria lysis occurred after induced with high temperature （42 ℃ ） , and the lysis efficiency of gene E - 15aalinker - SN was better than that of gene E. In the lysis process, the addition of Ca^2＋ and Mg^2＋ caused interference to the lysis effectiveness of protein E, while the addtion of Ca2 ＋ and Mg2 ＋ after inducing at 90 min could alleviate the interference. The experimental results showed no evidence of promotion effect of the addition of divalent cations on the formation of bacterial ghost.

关 键 词：大肠杆菌 菌影 双基因裂解系统 温控表达载体 裂解效果 

分 类 号：S852.612[农业科学—基础兽医学]