The Prokaryotic And Eukaryotic Expression Of The Hypoxia Inducible Factor-1 (HIF-1) From Taenia Solium

Taenia solium larvae, named Cysticercus cellulosae, can infect human and pig,usually in the tissues or organs of subcutaneous tissue, muscles, and brain. T. solium adult, as the cause agent of Taeniasis, parasitizes in the small intestinal of human body.T. solium adult is parasitic in hypoxia environment and low oxygen condition plays a vital role in the survival of the T. solium. The main roles of Hypoxia inducing factor-1(HIF-1) is to help the organization cell grow rapidly in low oxygen environment and promote glycolysis, etc. Therefore, in this study, T. solium HIF-1was cloned and expressed based on T. solium genome data base. The results are as follows:1. Total RNA was extracted from T. solium adult tissue and reversely transcribed into c DNA. Then T. solium HIF-1 was successfully cloned based on the c NDA template. The obtained ORF was 303 bp in length, encoding 100 amino acids, and the theoretical molecular weight of this HIF-1 protein was about 11.27 ku.2. The recombinant prokaryotic expression plasmid p ET-30a(+)-HIF-1 was constructed and inductively expressed. The recombinant protein was mainly expressed in the form of inclusion bodies in prokaryotic expression system. Western blotting analysis showed that HIF-1 protein purified was specifically recognized by the His labeled antibodies.3. Rabbits were immunized with the purified expression products of HIF-1.The serum was isolated to measure antibody titer after the third immunization. The antibody titer could reach to 1:51200.4. The recombinant eukaryotic expression plasmid pc DNA3.1-HIF-1 was constructed, which was then transfected into 293 T cell. Western blotting analysis showed that HIF-1 gene was successfully expressed in eukaryotic expression system.In conclusion, T. solium HIF-1 was successfully expressed in the prokaryotic and eukaryotic expression systems respectively. The recombinant protein could be recognized by the polyclonal antibodies. The polyclonal antibodies had high antibody titer. This laid the foundation to further research the functions and exact mechanisms of the HIF- 1 in T. solium.