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
Product Name pColiExpress™ I pColiExpress™ II pColiExpress™ III pColiExpress™ IV
Catalog Number BE001 BE005 BE010 BE015
Features
5'His6 Tag      
3'His6 Tag  
Periplasmic expression      
5'cmyc Tag      
HRV protease cut site      
Resistance Marker-Ampicillin
T7 promoter
Low copy number (ori pBR322)
Strain Protein
Expression
BL21 (DE3)
BL21 (DE3) pLys

  • T4 DNA Ligase based Bacterial Expression Kits
Product Name Applications
pColiExpress™ I Cloning & Expression Kit
  • Cloning of PCR fragments for subsequent expression of proteins in E. coli.
  • Expression of Proteins under the control of the T7 promoter.
  • Protein expression in BL21 (DE3) or BL21 (DE3) (pLys).
pColiExpress™ II Cloning & Expression Kit
pColiExpress™ III Cloning & Expression Kit
pColiExpress™ IV Cloning & Expression Kit

  • Competent Cells
Product Name Applications
BL21 (DE3) Competent Cells Protein Expression using the T7 Expression System.


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