The emergence of multidrug resistant bacteria in both community- and hospital-acquired infections is recognized as a major public health threat. Phage therapy is increasingly mediatized and researched as an additional tool for combatting antibiotic resistant infections. However, phages exhibit a number of properties that differ from antibiotics and hamper their development as pharmaceutical products and their application in therapy. This paper advocates a paradigm shift in the development and application of infectious disease therapeutics to cater for personalized phage therapy, which could be realized by the year 2035. More specifically, it presents a sustainable and ethical supply chain of instant synthetic phages, based on a community effort, supported and steered by public health organizations, and managed by a platform combining Artificial Intelligence (AI) and Distributed Ledger (DL) Technology.
Pascal Weibel, Miriam Ender, Jerzy Madon, Annelies S. Zinkernagel Ā· 5 authors
Introducing PCR products into plasmids vectors is key for molecular techniques. Ideally cloning vectors are easy to construct, modify and propagate, neither require advanced techniques nor special equipment or reagents and efficiently incorporate PCR products at close to zero empty vector background. We provide an easy to engineer self-made cloning vector, neither requiring sophisticated tools or techniques nor advanced cloning knowledge. Through recombination we obtained the pUC18ccdB vector, carrying the ccdB suicide gene within the pUC18 backbone. When SmaI cleaved (within the ccdB) vector was T4 ligated with small (0.2 kbp) and intermediate (1.3 to 2.2 kbp) blunt end PCR-products and transformed into E. coli, the amount of clones with incorporated PCR product was comparable to commercial PCR-cloning kits and at a close to zero PCR product negative background. In conclusion we present a simple, versatile and cheap approach to an efficient āhome made ā PCR-cloning vector that allows integration of crude blunt end PCR products at close to zero background.