Mutation Generation System Kit

Thermo Scientific transposon products are based on the transposition machinery of the bacteriophage Mu. During the lytic phase of the phage's life cycle the machinery replicates its genome by transposing repeatedly inside the host genome. The Mu transposition reaction has been modified into an in vitro reaction catalyzed by a single enzyme - MuA Transposase. In this system, one in vitro reaction is capable of generating more than a million transposon insertion clones.

The Mutation Generation System (MGS Kit) and Stop Generation System (STOP Kit) were developed for functional analysis of proteins. These new transposon tools enable the creation of saturated libraries of mutated proteins in a single reaction with less hands-on time than any other method. The location of the transposon insertion in each mutated clone can be mapped by either PCR or sequencing. With MGS and STOP kits, thousands of mutated clones are ready for expression studies in just 2 to 3 days.

The MGS Kit contains the complete set of reagents for transposon-based linker scanning mutagenesis of any target protein. The MGS Entranceposons are designed for making subtle changes in the structure of a target protein by inserting 15 bp in-frame linkers throughout the corresponding target gene. This in-frame insertion allows for conservation of downstream sequences.

The STOPKit Entranceposons contain translational stop codons in all three reading frames within the terminal portion of the transposon sequence. The proprietary modification of the Stop Generation System makes it possible to generate a saturated C-terminal deletion library from virtually any target protein with a maximum addition of three amino acids.

Features:

• Efficient—Create saturated insertion libraries for sequencing and protein analysis in a single reaction
• Fast—Decrease hands-on time compared to conventional methods
• Random—Eliminate target site preference or insertion hot-spot

Applications

The STOP Kit generates truncated proteins for functional assays of:

• Enzymes
• Receptors
• Structural proteins etc.

The MGS Kit generates random fifteen basepair in vitro insertions into any target DNA for:

• Rapid generation of in-frame five amino acid insertion libraries of any protein for functional analyses
• Rapid and random mutagenesis of cloned promoters and other regulatory DNA regions
• Random insertion of a NotI restriction enzyme site into any target DNA clone

Advantages
MGS Kit

• Thousands of different insertion clones from a single reaction
• Generates random insertions of 5 amino acids in all 3 reading frames
• Short in-frame insertions; no stop codons
• Flexibility in mapping mutants of interest: mutations are easily mapped by NotI or PCR
• Faster and more effective than linker scanning mutagenesis
• STOP Kit

• Saturated library of truncated proteins from a single reaction in two days
• Translational STOP codon in all three reading frames
• The target DNA sequence can be unknown
• Faster and more effective than conventional methods
• No specific primers required

Related Products
MuA Transposase
MuA Transposase (concentrated)