Towards the construction of high-quality mutagenesis libraries.
Biotechnol Lett
; 40(7): 1101-1107, 2018 Jul.
Article
em En
| MEDLINE
| ID: mdl-29700725
ABSTRACT
OBJECTIVES:
To improve the quality of mutagenesis libraries in directed evolution strategy.RESULTS:
In the process of library transformation, transformants which have been shown to take up more than one plasmid might constitute more than 20% of the constructed library, thereby extensively impairing the quality of the library. We propose a practical transformation method to prevent the occurrence of multiple-plasmid transformants while maintaining high transformation efficiency. A visual library model containing plasmids expressing different fluorescent proteins was used. Multiple-plasmid transformants can be reduced through optimizing plasmid DNA amount used for transformation based on the positive correlation between the occurrence frequency of multiple-plasmid transformants and the logarithmic ratio of plasmid molecules to competent cells.CONCLUSIONS:
This method provides a simple solution for a seemingly common but often neglected problem, and should be valuable for improving the quality of mutagenesis libraries to enhance the efficiency of directed evolution strategies.Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Biblioteca Gênica
/
Evolução Molecular Direcionada
Idioma:
En
Revista:
Biotechnol Lett
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
China