Combinatorial library approaches for improving soluble protein expression inEscherichia coli

Combinatorial library approaches for improving soluble protein expression in Escherichia coli.

High-throughput screening methodologies are already used in structural biology to define efficient protein crystallization and expression conditions. Recently, screening approaches have been extended to the optimization of genetic constructs for improved soluble protein expression. With similarities to the directed evolution strategies used in protein engineering, a target gene encoding a poorl...

cloning, expression and library construction for hiv-1 tat protein

background: designing novel therapeutic agents has been a critical challenge for hiv disease. materials and methods: in current study a dna sequence which was encoded the tat protein was synthesized and inserted in pet 28 vector. vector was cloned in bl21-de3 e. coli and cultured in tb media. after protein expression, recombinant tat protein was purified by nta affinity chromatography. the tat ...

BIOTECHNOLOGICALLY RELEVANT ENZYMES AND PROTEINS Expression of chondroitin-4-O-sulfotransferase inEscherichia coli and Pichia pastoris

Chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. While the enzymes involved in the biosynthesis of chondroitin sulfates are well known, the cloning e...

CoESPRIT: A Library-Based Construct Screening Method for Identification and Expression of Soluble Protein Complexes

Structural and biophysical studies of protein complexes require multi-milligram quantities of soluble material. Subunits are often unstable when expressed separately so co-expression strategies are commonly employed since in vivo complex formation can provide stabilising effects. Defining constructs for subunit co-expression experiments is difficult if the proteins are poorly understood. Even m...

Advances and challenges in recombinant protein expression in Escherichia coli