Cloning of human interleukin-18 binding protein and the construction of its retroviral vector

Objective To clone the cDNA of human interleukin-18 binding protein (hIL-18BP) and construct the recombined hIL-18BP retrovirus as a theoretical and preparatory approach to the IL-18BP gene therapy for autoimmune diseases. Methods The total RNA was extracted from the placenta of a Chinese individual and the intact cDNA of hIL-18BP was amplified by RT-PCR. The cDNA obtained was first linked with pcDNA3 vector and then transferred into E.coli DH5(. After being identified, the recombined plasmid (pcDNA3/ hIL-18BP) was used as the template in PCR, and finally, the 585 bp PCR product was inserted into the retroviral vector pLNCX. Results Recombined pcDNA3/ hIL-18BP, pLNCX/ hIL-18BP were identified by double-enzyme digestion and the results of sequencing were consistent with that of the published hIL-18BP cDNA. Conclusion Chinese IL-18BP cDNA has been cloned and the recombined hIL-18BP retroviral vector has been constructed successfully.