• Irshad Ahmad

• Sher Bahadar Khan

• Nighat Nawaz

• Mohammad Zahid Mustafa

• Asadullah

• Mutiullah Khattak

• Momin Khan

• Asif Ali

• Yasar Mehmood Yousafzai

• Sadeeq ur Rehman

• Chris Thompson

تلخیص

Transport protein GAB15256 from Arthrobacter globiformis belongs to the Major Facilitator Superfamily (MFS) family. In silico analysis of this protein is myo-inositol transporter but specific biological function is unknown yet. Therefore, large scale production and purification would help to study the exact role of GAB15256 protein. In this study, GAB15256 gene was cloned and over expressed in E. coli BL21 (DE3) strain. Membranes expressing GAB15256 were solubilised with 1% n-dodecyl-β-D-maltoside (DDM) and were purified by immobilised metal affinity chromatography using a Ni-NTA resin. Far UV circular dichroism spectroscopy demonstrates that purified protein GAB15256 was largely alpha helical secondary structure and has retained normal folding after passing through the various steps of purification. The thermal stability of GAB15256 was analysed by ramping the temperature from 5- 90 ºC and finally back to 5 ºC. On returning the temperature to 5 ºC, there was no evidence that suggests refolding of the protein and a melting temperature of 45.8 ºC was estimated using Global Analysis CD software 3.

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