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Updated: Jul 1, 2025

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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通过插入-删除策略,通过核酸酶的工程基质变质

Gaofei Liu1, Jialing Wang1, Jianlin Chu2

  • 1College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, China.

JACS Au
|March 1, 2024
PubMed
概括

工程核酸酸化酶表现出更广泛的基质特异性,使各种核酸类型的高效合成. 这一进步简化了以前难以制造的有价值化合物的生产.

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:
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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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