Production of recombinant enzymes of wide use for research|
Manzur, María J.; Muñoz, Rosana V.; Lucero, Adrián A.; Ayub, Maximiliano Juri; Alvarez, Sergio E. & Ciuffo, Gladys M.
For biotechnological purposes, protein expression refers to the directed synthesis of large amounts of desired proteins. The aim of the present work was to produce reverse transcriptase Moloney murine Leukaemia Virus retro-transcriptase and Taq DNA polymerase, as bioactive products. In the present paper, we report the preparation of recombinant enzymes, expressed in E. coli strains. The enzymes produced exhibited quite good activity, compared with commercial enzymes, allowing us to replace the last ones for several lab applications. We are reporting changes and modifications to standard protocols described. The standard protocols were modified, i.e. for the purification step of Taq, a temperature dependent procedure was designed. The enzymes produced were used in different applications, such as PCR, RT-PCR, PCR Multiplex and RAPDs molecular markers.
bioactivity, protein expression, purification, recombinant enzymes.