Thermo Scientific transposon products are based on the transposition mechanism of bacteriophage Mu. The device replicates the phage's genome by repeatedly transposing within the host genome during the lytic phase of its life cycle. This Mu transposition reaction has been modified into an in vitro reaction catalyzed by a single enzyme (MuA transposase). In this system, more than one million transposon insert clones can be generated using a single in vitro reaction.

Features

The Mutation Generation System (MGS Kit) and Stop Generation System (STOP Kit) were developed specifically for protein functional analysis. Using these new transposon tools, it is possible to create saturated libraries of mutant proteins in a single reaction, requiring less hands-on time than all other methods. By PCR or sequencing, the location of the transposon insertion in each mutant clone can be mapped. With MGS and STOP kits, thousands of mutant clones can be prepared for expression studies in just 2 to 3 days.

MGS kits contain a comprehensive set of reagents for transposon linker scanning mutagenesis of any protein of interest. MGS Entranceposons are designed to make subtle changes in target protein structure by inserting 15 bp in-frame linkers throughout the corresponding target gene. This in-frame insertion saves downstream sequence.

STOPKit Entranceposons Contains translation stop codons in all three reading frames of the terminal portion of the transposon sequence. The Stop Generation System's proprietary modification method allows you to construct saturated C-terminal deletion libraries from virtually any target protein, adding up to three amino acids

Features:

Efficient—in a single reaction Simultaneously construct saturated insert libraries for sequencing and protein analysis
Fast—shorter hands-on time compared to conventional methods
Random—eliminate target preference or insertion hotspots

Application

The STOP kit can construct truncated proteins for functional analysis of the following objects:

•Enzymes
• Receptors
• Structural proteins, etc.

MGS kit can insert randomly generated 15 base pairs into any target DNA in vitro for:

• Rapidly construct in-frame five amino acid inserts of any protein Libraries for functional analysis
• Rapid random mutagenesis to clone promoters and other regulatory DNA regions
• Randomly insert NotI restriction endonuclease sites into arbitrary target DNA clones

Advantages
MGS Kit

• Generate thousands of clones of different inserts in a single reaction
• Construct random insertions of 5 amino acids in all 3 reading frames Fragments
• Short in-frame insert; no stop codon
• Flexible mapping of targeted mutations: Easily map mutations via NotI or PCR
• Faster and more efficient than linker partitioning mutagenesis
• STOP Kit

• Constructs saturated libraries of truncated proteins in two days in a single reaction
• Contains translation stop codons in all three reading frames
• Target DNA sequence Can be unknown
• Faster and more effective than conventional methods
• No need for special primers

Related products
Mutation Generation System Kit
MuA Transposase