![]() Moreover, PCR-based methods are error prone and the restriction enzyme-based methods require specific recognition sequences to be present at specific sites and will in turn limit the number of fragments based on the number of restriction sites that can be used 6, 14. Some strategies, as designing adaptors that allow for sequences to be part of alternate libraries, only partially surpasses this limitation and in the process create scars and intermediary products are often incompatible with future assembling units 17. The homology-based methods require sequence overlap, which limit the type and order of fragment cloning. Inevitably, each method has its own disadvantages and so far, a platform capable of uniting flexibility, fidelity, efficiency and universality for unbiased handling of multiple DNA segments has yet to be developed. These methods are typically based on either sequence homology (e.g., isothermal assembly 12, recombination 13) or sequence signatures (also known as prefix and suffix) left by restriction digestion followed by ligation of DNA (e.g., BioBricks 14, GoldenGate 15) (for a review, see 16). Such studies and progresses however would never have been possible without the advances in the cornerstone of synthetic biology: DNA synthesis and assembly.Ī plethora of cloning methods is available for handling genes and/or gene parts, gene pathways and even subgenomes. These principles have been developed and deployed in several organisms 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11. ![]() ![]() Genetic circuits, in this context, consist of the systematic interactions between various molecular components (e.g., DNA activation/repression, RNA secondary structure, protein-dependent signaling, (in)organic molecules gradients) that are responsible for controlling and adjusting function and behavior in an organism. Synthetic biology encompasses conceptual design, construction, analysis, evaluation, tuning and remodeling of genetic circuits.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |