Deletion mutagenesis
A deletion mutagenesis ( English deletion mutagenesis ) is the genetic removal of sequences from the DNA by gene mutation . In contrast to insertion mutagenesis , a cloned DNA sequence is shortened in accordance with an excision or deletion . The sequence to be removed can be at the end of the sequence or within the DNA molecule.
execution
With the help of a restriction enzyme , the DNA sequence is cut at the ends of the DNA strand or in the middle of the strand. The thus produced DNA fragments may be with other fragments which complementary sticky ends (engl. Sticky ends ) or blunt ends (engl. Blunt ends ) have, using a ligase are linked.
If the process of deletion was carried out with different sequences and the different fragments formed are then joined together, a new DNA sequence with new properties is created. This can code for a gene expression (mostly protein ) of a new protein structure .
application
With the help of deletion mutagenesis, new proteins can be made from different domains . Unwanted properties or binding sites can be specifically removed so that, for example, side effects with biotechnologically produced proteins occur less.
For example, the hormone binding domain of a transcription factor can be coupled to another protein or removed by a deletion .
Individual evidence
- ^ H. Liu, JH Naismith: An efficient one-step site-directed deletion, insertion, single and multiple-site plasmid mutagenesis protocol. In: BMC biotechnology. Volume 8, 2008, p. 91, ISSN 1472-6750 . doi: 10.1186 / 1472-6750-8-91 . PMID 19055817 . PMC 2629768 (free full text).