Extraction of protease gene from recombinant host of Escherichia Colt BL21 (De3) Plyss

Thermostable protease are proven to be very effective in improving the industrial processes in many fields. As vast knowledge of creating new and improved ways to produce more production of thermostable enzymes it is now possible to express high production of enzymes using microbial technology with the correct methods and techniques. This is by altering the properties of the microbial protease and using recombinant technology and protein engineering obtain microbial protease that is ideal to be used in the biotechnology applications. The aim of this study is to extract the protease gene from the recombinant host, E. coli BL21 (DE3) pLysS. From the results, the host that has 50a protease gene was able to produce halo zones on the 10% skim milk agar after incubation at 60°C for 6 hours by which showed the ability of the protease enzyme to hydrolyse casein. The recombinant vector, pET was successfully extracted showed size of 3800 bp. Then the recombinant pET was used as a DNA template in order to amplify the protease 50a gene. The PCR product was successfully determined by 1% agarose and show the size of 1100bp. The amplified protease 50a gene then should be ligated to pGEX vector and transformed to other host for high expression of protease