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Genes and genomics
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2018
Polymerase cycling assembly is a method for the assembly of large DNA oligonucleotides from shorter fragments. This process uses the same technology as PCR but takes advantage of DNA hybridization and annealing as well as DNA polymerase to amplify a complete sequence of DNA in a precise order based on the single-stranded oligonucleotides used in the process. It thus allows for the production of synthetic genes and even entire synthetic genomes.
Design of artificial cells: artificial biochemical systems, their thermodynamics and kinetics properties
Published in Egyptian Journal of Basic and Applied Sciences, 2022
Adamu Yunusa Ugya, Lin Pohan, Qifeng Wang, Kamel Meguellati
An artificial cell should have the ability to self-replicate, get involved in a learning process, and keep homeostatic data (LYFE definition). In earlier concepts, von Neumann theory provides useful guidelines to build logically a self-replicating compartment [25]. However, to capture the entire spirit of cellular life, there is a need to consider many other nongenetic processes, e.g. molecular self-organization/crowding, artificial environment for nutrient exchange etc. Although the bottom-up approach motivates and also addresses the fundamental question of non-genetic processes, selective exchanges through phospholipid bilayer, osmotic pressure, transcription and translation processes are still challenging issues towards the development of workable microscopic vesicles. Recently, in a top-down approach [23], the genome of a living cell was knocked down to a minimum level to demonstrate that bacteria can be reprogrammed with synthetic genomes. Both bottom-up and top-down approaches towards the synthesis of artificial cells require the development of original methods and ideas [26]. Noireaux et al. demonstrated the states and the development of artificial cell synthesis with experimental constraints. Based on the basic idea of the development of a phospholipid-containing compartment encapsulating synthetic DNA, the expression of genes was carried in vitro upon the exchange of adequate nutrients. Moreover, cell-free transcription and translation permitted the expression of many genes. Nevertheless, the development and integration of synthetic DNA is still challenging for the synthesis of self-reproducing automatons and bacterial genomes [9] (Figure 1).