重组昆虫病原细菌Cip蛋白基因工程菌发酵条件的初步研究  

Preliminary studies on fermentation of recombinant crystalline inclusion protein of entomopathogeic bacterium in Escherichia coli

作　　者：游娟[1] 黄建林[2] 曹莉[3] 韩日畴[3] 

机构地区：[1]广东药学院生物化学与分子生物学系,广州51006 [2]广州市计量检测技术研究院,广州510030 [3]广东省昆虫研究所,广州510260

出　　处：《生物学杂志》2012年第1期25-29,共5页Journal of Biology

基　　金：广东药学院基金(2006JCX07)

摘　　要：昆虫病原细菌产生两种胞内晶体蛋白CipA和CipB,为新型的生物农药——昆虫病原线虫提供必需的营养。在已构建原核表达载体pET-15b-cipA和pET-15b-cipB的基础上,研究了这两种重组载体在不同宿主菌中的表达情况,重组菌的生长特性,摇瓶培养时表达重组Cip蛋白工程菌的发酵条件。结果显示:以E.coli BL21(DE3)为宿主菌,在LB培养基中培养至OD600为0.8～1.0时,1 mmol/L IPTG诱导8 h,CipA和CipB的表达量可达30.9%和32.6%,重组质粒具有良好的分离稳定性和结构稳定性。Two types of intracellular crystalline inclusion proteins produced by Photorhabdus bacteria named CipA and CipB are the key nutrient source for the novel biopesticide entomopathogeic Heterorhabditis nematodes. Based on constructed pET-15b-cipA and pET- 15b-cipB, the protocaryotic expression and fermentation of recombinant Cip proteins were studied. When recombinant E. coli BL21 （ DE3 ） was induced in LB medium with 1 mmol/L IPTG added at OD600 0. 8 - 1.0 for 8 hours, CipA and CipB expression constituted 30. 9% and 32. 6% of the total bacterial protein, respectively. Both recombinant strains showed stable plasmid inheritance and consistence. This research will be helpfnl for the fermentation of Cip on a large scale and promote the commercialization of nematode products.

关 键 词：胞内晶体蛋白 基因工程菌 发酵 

分 类 号：Q78[生物学—分子生物学]