Abstract :
Lactobacillus plantarum is a lactic acid bacterium, mostly found in fruits and vegetables. It has been used in a variety of food fermentations. It is reported that strains from this species have probiotic activity. In this study, pUC19 was chosen as a vector to clone ?-galactosidase gene. To clone this gene we used EcoRI and KpnI restriction enzymes that give a DNA fragment of 5000 bp in length which contains ?-galactosidase gene. The same endonuclease enzymes were used to cut the vector (pUC19). To know the length of isolated DNA fragment as well as digested pUC19, agarose gel electrophoresis was used along with Lambda HindIII and Hyperladder I respectively. QIAGEN kit was used to extract DNA fragments from the agarose gel. Then it utilised for ligation in further processes. Furthermore, the DNA fragments were transformed into host cells (E. coli) and they were spread on LB agar plates containing X-gal and IPTG to confirm the presence of inserted DNA.
Keyword :
Lactobacillus plantarum, ?-galactosidase, Probiotics, Cloning.