pColiExpress™ Bacterial Expression

The traditional methods of inserting genes into vectors are based on DNA cleavage by restriction endonuclease and then ligation by DNA ligase. However, this approach to DNA engineering is time-consuming and relatively inefficient. Ligation Independent Cloning (LIC) technique is developed as an alternative to restriction endonuclease/ligase cloning. LIC provides a simple and cost-effective tool for producing many constructs of a single target or multiple targets in parallel without the need to select specific restriction enzymes for each gene.

T4 DNA polymerase is a template-dependent DNA polymerase that has 3’-exodeoxyribonuclease activity, but lacks 5’ to 3’ exodeoxyribonuclease activity. LIC uses the 3´-exodeoxyribonuclease activity of T4 DNA polymerase to assemble DNA molecules by non-covalent complementation of single-stranded DNA of the insert and vector. Briefly, LIC relies on about 10- to 15-nucleotide complementary 3′ overhangs at the ends of a PCR-amplified DNA fragment and a linearized vector to make a stable hybridization product that can be readily used to transform host organisms without ligation. LIC based on T4 DNA polymerase cloning is strictly sequence-dependent as it requires the presence or absence of specific nucleotides at certain locations in the overlapping region.

pColiExpress™

pColiExpress™ LIC Cloning & Expression Kits are a highly efficient, versatile and fast system of DNA cloning vectors for protein expression in E. coli. All family systems are based in directional LIC technology, a rapid procedure that provides high-cloning efficiency and the fast production of a large quantity of any desired proteins.

Advantages & Features

• Ready-to-use vectors.
• Highly efficient Cloning System with Higher Protein Expression levels.
• Special design that allows the Cell to keep larger numbers of copies than other plasmids with ori pBR322.
• Linearized vector: ready for ligation with your PCR amplified with the recommended Primers.
• Minimum background: lower than 1%.
• Time-saving protocol: avoids any step required after PCR.
• Cost avoidance: avoids the use of expensive Phosphorylated Primers.
• Versatility: cloning of PCR fragments amplified with any type of DNA Polymerase.
• Risk-free: product covered by our Quality 100% Guarantee.

Product Range

• LIC technology based Bacterial Expression Kits

• T4 DNA Ligase based Bacterial Expression Kits

• Competent Cells