CLONING AND EXPRESSION OF INVERTASE GENE FROM INVERTASE PRODUCING BACTERIA TO NON-PRODUCING BACTERIA

Abstract

In the present study, the gene of invertase enzyme from Bacillus megaterium was cloned and expressed in non-invertase producing E. coli. Bacillus megaterium was isolated from soil and identified by performing various biochemical tests. Then the isolation of genomic DNA followed by electrophoresis on a 0.8% agarose gel was done. Amplification of gene of interest using invertase primer was done by PCR. The amplified product observed in 1.2% gel stained with ethidium bromide. This PCR product was further subjected to restriction digestion (EcoRI) and then ligation with the vector (Plasmid DNA pUC18). A competent cell of the target bacteria (invertase non-producing) was prepared using chemical method (CaCl₂). The ligation mixture was allowed to react with the competent cells in order to uptake the DNA insert ligated with the vector. The mixture was spread over the nutrient agar plates added with the antibiotics (Ampicillin). The transformed E. coli cells were screened by observing the presence of colonies over the nutrient agar plate as the transformed cells showed the property of antibiotic resistance.