pbactag GFP+ plasmid vector枯草芽孢杆菌荧光表达质粒载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

pbactag GFP+ plasmid vector枯草芽孢杆菌荧光表达质粒载体

The GFP+ tag (pBacTag-GFP+) is an improved variant of the common GFP, which produces enhanced fluorescence.
The mechanism of pBacTag Tagging Vectors is illustrated with pBacTag-GFP+ as example. This vector can be used for creating a GFP+ fusion protein from any chromosomally located gene of interest, by fusing a gfp+ tag to the chosen gene. In this example the gene of interest is named orf2. It is part of an operon, including three genes in total (orf1, orf2 and orf3). For getting the gfp+ tag fused to the orf2, the 3’ part of the gene (orf2’) has to be inserted into the multiple cloning site of the pBacTag-GFP+ vector. After transforming B. subtilis cells with this construct, chromosomal integration of the vector is achieved by selecting for cells with resistance against erythromycin. The integration is facilitated by homologous recombination of both orf2’ copies (one copy being within the plasmid, the other one within the chromosomal DNA). The mechanism the pBacTag-GFP+ vector (with orf2’) is integrated into the genome

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