中文摘要：

以干酪乳杆菌（Lactobacillus casei）组成型表达质粒p PG-T7g10-PPαT为重组载体,在限制性内切酶SacⅠ和ApaⅠ酶切位点之间,插入增强型绿色荧光蛋白（EGFP）基因,构建重组质粒p PG-T7g10-EGFP,电转化干酪乳杆菌ATCC393（L.casei 393）,获得重组菌p PG-T7g10-EGFP/L.casei 393;利用Western blot检测可见约27 ku的重组蛋白特异性反应带;激光共聚焦显微镜下可见重组菌出现特异的绿色荧光;流式细胞术分析可见重组菌出现了明显的荧光信号。结果表明：已经获得组成型表达EGFP的重组干酪乳杆菌。本研究为进一步利用该表达系统表达功能蛋白,拓展乳酸菌的应用提供了参考。

英文摘要：

Enhanced green fluorescent protein（ EGFP） gene was inserted into Lactobacillus casei constitutive expression plasmid p PG-T7g10- PPαT using the restriction enzymes SacⅠ and ApaⅠ and thus construct a recombinant plasmid p PG- T7g10- EGFP. The recombinant vector was transformed into L. casei ATCC393 by electroporation to obtain the recombinant strain p PG- T7g10- EGFP / L. casei 393. The result of Western blot showed that a protein band with the molecular weight of 27 ku was detected. Green fluorescence could be observed by confocal laser microscope in the recombinant strain. Further flow cytometry analysis showed a clear fluorescent signal in the recombinant strain. These results suggest that a recombinant L. casei with constitutive EGFP expression has been obtained. This study will provide a basis for utilization of the functional protein expression system.