Cloning a synthetic gene for human stefin B and its expression in E. coli.

A gene coding for human stefin B was synthesized by the solid-phase phosphite method and cloned in the pUC8 cloning vector. The insert with the verified DNA sequence was subcloned into two expression vectors and expressed in E. coli as a fusion protein with beta-galactosidase and as a native protein.

Regen: program for designing gene assembly.

One of the main problems in constructing synthetic genes is the incorrect hybridisation between the oligonucleotides. The problem is resolved if the sequence uniquely defines the position of the oligonucleotide in the assembled gene. This can be accomplished through the wise partition of dsDNA sequence in the fragments.