Random Insertion and Deletion Mutagenesis

2003 ◽  
pp. 53-64 ◽  
Author(s):  
Hiroshi Murakami ◽  
Takahiro Hohsaka ◽  
Masahiko Sisido
Keyword(s):  
Deletion Mutagenesis ◽  
Insertion And Deletion ◽  
Random Insertion

scholarly journals In vitro evolution of antibody affinity via insertional scanning mutagenesis of an entire antibody variable region

2020 ◽  
Vol 117 (44) ◽  
pp. 27307-27318
Author(s):  
Kalliopi Skamaki ◽  
Stephane Emond ◽  
Matthieu Chodorge ◽  
John Andrews ◽  
D. Gareth Rees ◽  
...  
Keyword(s):  
Point Mutations ◽  
Variable Region ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Antibody Affinity ◽  
In Vitro Evolution ◽  
Deletion Mutagenesis ◽  
Insertion And Deletion ◽  
Random Insertion

We report a systematic combinatorial exploration of affinity enhancement of antibodies by insertions and deletions (InDels). Transposon-based introduction of InDels via the method TRIAD (transposition-based random insertion and deletion mutagenesis) was used to generate large libraries with random in-frame InDels across the entire single-chain variable fragment gene that were further recombined and screened by ribosome display. Knowledge of potential insertion points from TRIAD libraries formed the basis of exploration of length and sequence diversity of novel insertions by insertional-scanning mutagenesis (InScaM). An overall 256-fold affinity improvement of an anti–IL-13 antibody BAK1 as a result of InDel mutagenesis and combination with known point mutations validates this approach, and suggests that the results of this InDel mutagenesis and conventional exploration of point mutations can synergize to generate antibodies with higher affinity.

scholarly journals Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli.

1985 ◽  
Vol 161 (3) ◽  
pp. 1219-1221 ◽  
Author(s):  
S C Winans ◽  
S J Elledge ◽  
J H Krueger ◽  
G C Walker
Keyword(s):  
Escherichia Coli ◽  
Deletion Mutagenesis ◽  
Insertion And Deletion

The use of a drug resistance cartridge forin vitro insertion and deletion mutagenesis of a cosmid clone

Plasmid ◽  
1990 ◽  
Vol 24 (2) ◽  
pp. 149-152 ◽  
Author(s):  
Xing-Mai Jiang ◽  
Lajwant K. Romana ◽  
Peter Reeves
Keyword(s):  
Drug Resistance ◽  
Cosmid Clone ◽  
Deletion Mutagenesis ◽  
Insertion And Deletion
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