Thermococcus gorgonius DNA Polimerazının Rekombinant Olarak Üretimi

Year 2017, Volume: 43 Issue: 1, 1 - 9, 28.04.2017

Bilge Hilal Çadırcı , Oğuzhan Bebek Deniz Çam

Abstract

Termostabil bir enzim olan Tgo DNA polimeraz
hipertermofilik bir arkea olan Thermococcus
gorgonarius tarafından sentezlenmektedir. Çalışmamızda 3’-5’ ekzonükleaz
aktivitesiyle yüksek kalitede DNA sentezi gerçekleştiren Tgo DNA polimeraz,
rekombinant olarak üretimi hedeflenmiştir. Bu amaçla, ürettirilen yapay gen
uygun restriksiyon enzimleri ile kesilip aynı enzimlerle kesilmiş olan pET28a
vektör sistemine ligasyon ile birleştirilmiştir. E. coli BL21 hücrelerinde eksprese edilen enzim aktivitesi, PCR
reaksiyonu gerçekleştirilerek kontrol edilmiştir.

Keywords

DNA polimeraz, PCR, Rekombinant DNA teknolojisi, Thermococcus gorgonarius

Recombinant DNA Polymerase Production of Thermococcus gorgonarius

Abstract

A thermostable enzyme Tgo DNA polymerase was produced by a hyperthermophilic archaea, Thermococcus gorgonarius. In this study, it was aimed to produce the Tgo DNA polymerase which synthesizes highly qualified DNA by its 3’-5’-exonuclease activity, by recombinant DNA technology. For this purpose, synthesized synthetic DNA was cut with restriction enzymes and ligated into a pET28a vector. The activity of the enzyme expressed in E. coli BL21 was controlled by PCR reaction.

Keywords

DNA polymerase, PCR, Recombinant DNA technology, Thermococcus gorgonarius

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