Cloning of ?-Amylase enzyme producing gene into non?-amylase producing organism and its Expression

International Journal of Biotech Trends and Technology (IJBTT)
© 2014 by IJBTT Journal
Volume - 4 Issue - 1                          
Year of Publication : 2014
Authors :Parvati Saiyam , Kuldeep Diwedi , Sujeet K. Singh , Shruti Shukla , Madhulika Singh


"Cloning of ?-Amylase enzyme producing gene into non?-amylase producing organism and its Expression",International Journal of Biotech Trends and Technology (IJBTT), V4(1):1-3 Jan - Mar 2014. Published by Seventh Sense Research Group.


In this work, the gene of ?-amylase from B. subtilis was cloned and expressed in Non-amylase produsing E.coli. B. subtilis was isolated by performing various biochemical tests. The organism producing ?-amylase enzyme was successfully isolated. The isolation of genomic DNA followed by amplification of Gene Of Interest using ?-amylase primer and annealing temperature 56°C was done using PCR technique. The amplified product was eluted from electrophoresis gel and subjected to restriction digestion (EcoRI) and then ligation with the vector (Plasmid DNA pUC18). A competent cell was prepared using chemical method (CaCl2). The ligation mixture was allowed to react with the competent cells in order to produce transformed cells producing ?-amylase enzyme. The mixture was spread over the starch plates. The successfully transformed E.coli cells were observed over the Starch agar plate.


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Bacillus subtilis, E.coli, PCR, pUC18, EcoRI, etc.