Cinquin Lab

Menz R.I., Cinquin O., Christopherson R.I. Ann. Trop. Med. Parasitol. 96(5), pp469-476 (2002)

Abstract

The coding region of a putative orotidine 5′-monophosphate decarboxylase gene from Plasmodium falciparum was identified in genomic data from the Malarial Genome Sequencing Project. The gene encodes a protein of 323 amino acids with a predicted molecular weight of 37.8 kDa. The gene was cloned into a bacterial expression vector and over-expressed in Escherichia coli. The recombinant protein was purified and shown to have orotidine 5′-monophosphate decarboxylase activity, confirming the identity of the gene.

Cinquin O., Christopherson R.I., Menz R.I., Mol. Biochem. Parasitol. 117(2), pp245-247 (2001)

Abstract

The RIG plasmid [Baca and Hol, 2000, Int. J. Parasitol. 30, 113-118] encodes tRNA genes rare in Escherichia coli, enabling codon-biased Plasmodium genes to be over-expressed in E. coli. Expression of orotidine-5′-monophosphate decarboxylase (EC 4.1.1.23) from Plasmodium falciparum was found to be toxic to E. coli BL21(DE3) cells carrying the RIG plasmid. Unfortunately, the RIG plasmid is incompatible with the pLysS plasmid commonly used with pET expression vectors to allow over-expression of recombinant proteins toxic to E. coli by repressing their basal level expression. We describe here a new hybrid plasmid carrying the gene for T7 lysozyme and those for the rare E. coli codons argU, ileX and glyT, which facilitates over-expression of orotidine-5′-monophosphate decarboxylase (EC 4.1.1.23) from P. falciparum.