[Cloning and expression of triosephosphate isomerase gene of Boophilus microplus]

Total RNA was extracted from adult Boophilus microplus. RT-PCR was used to amplify the gene and the fragment was subcloned into the expression vector pET-28a. The cloned gene was expressed in E. coli Rosseta (DE3), induced by IPTG, and identified by SDS-PAGE. The results showed that the triosephosphate isomerase (tim) gene of B. microplus has 750bp and encodes 249 amino acids (GenBank No. JX112888). The cloned tim gene shares 99% homology with that in tick embryos. The relative molecular weight (M(r)) of the expressed recombinant protein is about 27 000.