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Öğe Synthetically engineered Bacillus subtilis inhibits growth of Escherichia coli by secreting Limulus polyphemus anti-lipopolysaccharide factor(Canadian Soc Clinical Investigation, 2015) Guver, Alperen; Gunbey, Betul; Yilmaz, Fazilet G.; Gungordu, Halenur; Soylu, Hasan Y.; Elitok, Mustafa S.; Bayrak, Ozan S.Purpose: E. coli is a gram-negative bacterium. Outbreaks of the enterohemorrhagic E. coli strain (EHEC) have caused dangerous pandemics and dozens of deaths. The lipopolysaccharide (LPS) endotoxin cell wall of gram-negative bacteria is an important cause of septic shock. The aim of this study was to design an innovative genetic construct model whose expression prevents gram-negative bacteria growth and infection. Noninfectious gram-positive bacteria B. subtilis were modified with a constitutive secretory gene construct to secrete Limulus polyphemus anti-lipopolysaccharide factor (LALF), which binds and neutralizes the LPS layer found in E. coli cell wall. Methods: LALF gene construct was designed by using the Gibson assembly synthetic biology cloning method for the purpose of synthesis and secretion of the protein in B. subtilis. The synthesis and secretion of LALF protein was optimized with constitutive promoter, ribosome binding site and secretory signal peptide specific for B. subtilis gene expression system. Results: In the zones on LB-agar growth plate where LALF protein-secreting B. subtilis supernatant had been added, growth of red fluorescent protein (RFP) expressing E. coli was inhibited. There was no E. coli colony on the zones where LALF secreted B. subtilis biofilm has been added. Conclusion: LALF inhibits the growth of gram-negative bacteria. LALF may prove to be useful for the treatment of diseases caused by gram-negative bacteria. This study presents not only an alternative sterilization method for hospitals and laboratories but also an effective clearance treatment for bacteria that cause diseases such as septic shock.Öğe Tissue Engineering: Towards Development of Regenerative and Transplant Medicine(Academic Press Ltd-Elsevier Science Ltd, 2018) Elitok, Mustafa S.; Gunduz, Esra; Gurses, Hacer E.; Gunduz, Mehmet[Abstract Not Available]
