Construction of recombinant Lactobacillus expressing green fluorescent protein gene and transformation conditions by electroporation
TIAN Xing-shan~1, ZHANG Ling-hua~(1,2), ZHOU Feng-zhen~1, LI Guo-li~1, SUN Xiao-gang~1(1 Bio-Tech Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China; 2 College of Biological Engineering, South China University of Technology, Guangzhou 510640, China)
A recombinant Lactobacillus expressing green fluorescent protein (GFP) gene was constructed using recombinant DNA techniques. The GFP gene was excised from plasmid pGFP2 and subcloned into expression vector pThioHisB as an NcoⅠ/SacⅠ fragment.The resulting recombinant plasmid was named as pGhisGFP and then transformed into the BL21 strain of E. coli. The recombinant plasmid pThioGFP was extracted from the BL21, and then transformed into Lactobacillus by electroporation. The optimum parameters for electroporation were:D_(600 nm) 0.6, voltage 2.2 kV, buffer PEB, 100 μL cell volume.The recombinant bacterial grew on LB plate as green colonies, and the results showed that the GFP gene was expressed stably in the host bacterial (Lac1001).
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