Molecular cloning and expression of the specific region of the gene encoding the 83 kD protoplasmic cylinder protein from Borrelia burgdorferi B31 strain.

Aim\ To provide the target antigen for the development of a Lyme disease vaccine and serodiagnosis reagent.Methods\ According to the published gene seqence encoding the 83kd protoplasmic cylinder protein (p83) from Borrelia burgdorferi B31 strain, we selected the region from 1420bp to 1896 bp of the p83 gene as the target sequence to design a pair of oligonucleotide primers, obtained the gene fragment by using polymerase chain reaction, and constructed the recombinant plasmid by cloning the gene fragment into plasmid vector pBK-CMV. The recombinant plasmid was identified by restriction enzyme analysis and PCR amplification and DNA sequencing. Then, the recombinant plasmid was tranformed into E.coli XL-1 Blue MRF' and induced with IPTG. The expression product was analyzed by using SDS-PAGE and western-blotting. Results\ 1) The gene fragment encoding the specific region of the 83kD protoplasmic cylinder protein from Borrelia burgdorferi B31 strain was successfully amplified. 2) The purified PCR product was cloned into the BamHI and EcoRI site of the plasmid vector pBK-CMV, and a recombinant plasmid pBX1 was obtained. 3) Plasmid pBX1 can stably express a 29kD recombinant fusion protein in E.coli XL-1Bue MRF'. Conclusion\ A recombinant plasmid which contains the gene fragment encoding the specific region of the 83kD protoplasmic cylinder protein from Borrelia burgdorferi was successfully constructed. The recombinant plasmid can stably express a 29kD recombinant fusion protein in E.coli XL-1 Blue MRF'. These results provided the basis for further study the usefullness of the fusion protein in serodiagnosis and vaccine for Lyme disease.