Construcation of Vector for iclR knockout in P Solubilizing Klebsiella Pneumoniae SM6 and SM11

Abstract

In the present study, GFP gene was incorporated in pUC18 cloning vector for the selection and tagging of fluorescent protein. The isolated plasmids pUC18 and pRV85 were digested with the help of EcoRI. The GFP gene was present in between the EcoRI site in pRV85. Thus, pRV85 was digested with EcoRI. Digestion yielded a GFP fragment of 1000 base pairs which was observed on agarose gel. Subsequently, the GFP gene fragment was extracted from pRV85 and was ligated into the cloning vector, pUC18 (2.7 kb). The resulting pUC18-GFP was around a fragment of 3.7kb. For the confirmation of cloning, the pUC18-GFP was digested with EcoRI, where we observed two bands (2.7 kb and 1000 base pairs). Further, transformation of GFP into transformation host E. coli DH5α was performed. The GFP gene was successfully cloned and transformed into E. coli DH5α and fluorescent colonies were selected. From this study, it was concluded that the green florescent protein tagging was useful for detecting the bacteria in labelled fluorescent in vitro as well in vivo.