High-Level Production of MMLV Reverse Transcriptase Enzyme in Escherichia Coli

Reverse transcriptase (RT) of Moloney murine leukemia virus (MMLV) is the most widely used enzyme for cDNA synthesis and RNA amplification. In this study, we aimed to produce MMLV RT recombinantly due its importance in molecular studies. context, DNA fragment encoding was cloned into pTOLT plasmid expressed E. coli BL21 (DE3) pLysE cells. Since high-level expression protein caused molecules aggregate inclusion bodies, co-expression chaperone plasmids (pG-KJE8, pGro7, pKJE7, pGTf2, pTf16) performed obtain soluble form. Contrary our expectations, because it could not be obtained form, recovered from bodies using refolding process. Finally, purified by affinity chromatography activity checked RT-PCR technique.