重组UK114工程菌的高密度发酵培养  

High Cell Density Culture of the Engineering Bacterial Strain of the Recombinant UK114

作　　者：尹淑琴[1] 常泓[1] 范艳[1] 朱宏[1] 梁娟[1] 

机构地区：[1]山西农业大学生命科学学院,山西太谷030801

出　　处：《山西农业科学》2015年第8期924-926,931,共4页Journal of Shanxi Agricultural Sciences

基　　金：山西省科技攻关项目(20090311037)

摘　　要：为了鉴定重组UK114基因工程菌是否稳定表达,并进行初步的高细胞密度发酵试验。将重组基因工程菌BL21(DE3)/p GEX-4T-3-UK114进行菌种的传代培养并酶切检测;在保持菌种不变情况下,在5 L高密度发酵罐中,采用补料分批培养方法进行高密度发酵。结果表明,重组基因工程菌BL21(DE3)/p GEX-4T-3-UK114传代20代后质粒稳定存在,高密度发酵结束菌体浓度OD600大于50,融合蛋白量占菌体总蛋白量的31%,其含量达到2.1 g/L,工程菌获高效表达。To construct the engineering bacterial strain of the recombinant UK 114 for the high effective expression, the recombinant vector pGEX-dT-3-UK114 was constructed, and transformed into E. coli BL21 （DE3）. The expression of fusion protein GST - UK114 was induced with IPTG. The recombinant prokaryotic expression vector was successfully constructed by the restriction enzymes digestion and gene sequencing. Fed-batch culture was used to obtain high density culture cell and produce GST-UK114 fusion protein by 5 L fer- mentor. Fusion protein GST-UK114 was expressed in solubility in E. coli BL21 （DE3）in SDS-PAGE gel. The final density of grown bacterial was 51 （OD~）, the amount of fusion protein accounted for 31% of the total protein; the products of GST-UK114 was 2.1 g/L. The engineering bacterial strain of the recombinant UK1 lg （BL21 （DE3）/pGEX--4T-3-UK1 lg） was successfully constructed and ex- pressed in prokaryotes by high cell density culture with high efficiency. The results provide a foundation for studing the biological activity of UK 114 protein.

关 键 词：UK114 高密度发酵 GST融合蛋白 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]