Protein Extraction

Total leaf protein from transgenic and control tobacco plant was extracted in extraction

buffer. Crude extract of transgenic and control tobacco leaves was obtained by grinding 1g of

leaves in 3 mL of extraction buffer (0.2 M phosphate buffer, pH 7.0). The leaves were processed

in pre-cooled mortar and pestle. The homogenate was taken in eppendorf tubes and centrifuged at

20,000 g for 20 minutes in refrigerated centrifuge at 4 C. The supernatant was transferred to a

new sterile 1.5 mL ependorff tube for further use and stored at 4C.