Cloning of The Gene Encoding the Enolase from Acinetobacter baumannii


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Ata A., Uğurel E., Uğurel O. M. , İnci T. G. , Balık D.

Eurasian Congress on Molecular Biotechnology (ECOMB2019), Trabzon, Türkiye, 19 - 21 Eylül 2019, ss.109

  • Basıldığı Şehir: Trabzon
  • Basıldığı Ülke: Türkiye
  • Sayfa Sayıları: ss.109

Özet

Acinetobacter baumannii is a gram-negative, obligate aerobic, non-motile, oppurtunistic human pathogen that causes hospital-acquired infections, including pneumonia and blood-stream infections. The first ever list of antibiotic resistant “priority pathogens” was published by WHO. A. baumannii was being chosen as priority 1 (critical), emphasising its critical threats to public health because of its tendency to acquire multidrug, extensive drug and even pandrug resistance phenotypes. This list shows that new alternative drug candidates should be developed for treating A. baumannii infections. Structure-based drug design approaches can be studied for developing new alternative drug candidates. At the first step of structure-based drug design, in vitro analyses such as identification, cloning, expression and purification of target molecules are carried out. Enolase was selected as target  molecule for structure-based drug design approach. Enolase is a key glycolytic enzyme which catalyzes the dehydratation of 2-phosphoglycerate to phosphoenolpyruvate in the cytoplasm of prokaryotic and eukaryotic cells. In this research, genomic DNA from A. baumannii was isolated by using boiling method. Forward and reverse primers were designed according to nucleotide sequence of enolase from A. baumannii strain AB030. The gene was amplified by PCR. The amplified gene was inserted into the pGEM-T Easy vector and transformed into Escherichia coli DH5α. The gene encoding enolase from A. baumannii was cloned for the first time in this research.

Key words: Acinetobacter baumannii, Antibiotic Resistance, Enolase, Structure-Based Drug Design

Acknowledgments: This research has been supported by Yıldız Technical University Scientific Research Projects Coordination Department. Project Number: FYL-2019- 3496