Derivatives of the bacterial plasmid pET-16b containing genes from bacteriophage T4 that allow for the expression of one DNA (T4Dnl) and two RNA ligases (T4Rnl1A, and T4Rnl2) have been developed.
- Allow over-expression of recombinant versions of three ligases
- Ability to ligate a range of DNA-RNA hybrids
- Can be incorporated into Escherichia coli
- Contain a deca-histine tag at the N-terminus for easy purification
Ligases are vital for the production of recombinant DNA used to insert genes into plasmids, which are important tools in genetic engineering where they are used to express, clone or amplify certain genes. Ligases are, therefore, commercially-valuable proteins. Ligases have many other uses, such as being used to isotopically label RNA fragments by ligating labelled and unlabelled fragments together for use in NMR experiments to study the secondary structures present. There is an ever-increasing demand for more efficient and diverse ligases.
Many biotechnological companies investigate ligases due to their importance in the regulation of gene expression. The optimised enzyme activities produced from this project could improve biotechnological tools that are targeted at research into novel small RNAs.
Biochem. J., 2006, 398, 135-144. doi:10.1042/BJ20060313.
Team led by Dr Richard Bowater
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