2015年10月7日 · Importantly, MP6 can enable ∼ 2.3 substitutions per kbp in a gene of interest using phage vectors in a single generation, with additional increases in mutagenesis efficiency concomitant with ...

2014年2月2日 · In E. coli, full induction of the most mutagenetic MP (MP6) results in a ~300,000-fold increase in the mutation rate of chromosomal DNA (Table 1), which corresponds to ∼ 2.3 substitutions per kb...

Plasmid MP6 from Dr. David Liu's lab contains the insert dnaQ926 dam seqA emrR ugi CDA1 and is published in Nat Commun. 2015 Oct 7;6:8425. doi: 10.1038/ncomms9425. This plasmid is available through Addgene.

2024年3月15日 · The most effective MP, MP6, featured six genes that impede proofreading, mismatch repair, base excision repair, and base selection while increasing cytidine deaminations. MP6 was used to evolve a complex metabolic pathway consisting of ten genes (15.4 kb), demonstrating its applicability for metabolic engineering in E. coli [ 54 ].

在该Protocol中，作者首先总结了近年来其开发的各种突变率提高的突变质粒，最新版突变质粒 MP6 可以在10h内完成第一版MP1 200h的工作。 在定向进化中，选择严格性既影响进化的可行性，也影响进化蛋白的活性。

2023年8月31日 · Simultaneous expression of mutagenic proteins from the MP6 plasmid mutagenizes the phage, including the gene of interest. 67 During PACE, continuous dilution of a fixed-volume ‘lagoon’ with fresh host cells selects for rapidly propagating phage encoding molecules that trigger gIII expression (Figure S2 A).

2020年5月22日 · The most potent of these plasmids, mutagenesis plasmid 6 (MP6), encodes a cassette of six mutagenic genes affecting proofreading, mismatch repair, base selection, and base excision.

2020年10月16日 · Currently, the most effective variant of the MP is MP6 (available from Addgene), which increases the mutagenesis rate to more than 300 000-fold over the basal level of mutation in E. coli. This high mutagenesis rate allows for the rapid accumulation of mutations that persist in the protein after arabinose induction of MP. In theory, the PACE ...

2020年11月1日 · A mutagenesis plasmid, MP6, was then used to increase the mutation rate. In phage-assisted continuous evolution (PACE), the defective type of M13 with PD1 was used to infect the host bacteria and express PD1, which the expressed PDL1 of accessory plasmid within the host bacteria could then interact with.

Phage-assisted continuous evolution (PACE) enables continuous directed evolution in bacteria by mapping the steps of Darwinian evolution onto the bacteriophage life cycle and allows directed evolution to occur on much faster timescales compared to conventional methods.