Recombinant Echinococcus multi-Bb-Em14-3-3 Construction and expression efficiency of the vaccine.doc

 PAGE \* MERGEFORMAT 17 Recombinant Echinococcus multi-Bb-Em14-3-3 Construction and expression efficiency of the vaccine [Abstract] Objective To construct a multi-Echinococcus (Em) re-Bifidobacterium (Bb)-Em14-3-3 vaccines, and research Em14-3-3 molecule in E. coli BL21 (DE3) Expression efficiency. PCR amplification methods by Em14-3-3 antigen gene, the gene was cloned containing glutathione-S-transferase (GST) gene of Escherichia coli - Bifidobacterium shuttle vector pGEX-1T, The recombinant plasmid pGEX-Em14-3-3, electroporated the plasmid into Bb, build more recombinant Echinococcus Bb-Em14-3-3 vaccine. by PCR and enzyme digestion after IPTG induction Em14-expression 3-3/GST fusion proteins, SDSand the Western blot identification of the expressed product. Results PCR successfully amplified 530 bp of Em14-3-3 gene, restriction analysis confirmed that Em14-3-3 gene was inserted into pGEX-1T in, PCR confirmed that recombinant Bb-Em14-3-3 vaccine constructed successfully, SDSand Western blot analysis showed that recombinant plasmid was transformed into host bacteria highly expressed in the IPTG induced a Em14-3-3/GST fusion protein expression efficiency was 23 %. Conclusion We successfully constructed recombinant Echinococcus multi-Bb-Em14-3-3 vaccine, the recombinant plasmid pGEX-Em14-3-3 was highly in E. coli expression, Western blot results suggest that expression out of Em14-3- 3 recombinant protein with a specific antigen. [Keywords:] multiple Echinococcus, recombinant Bb-Em14-3-3 vaccine, construction, expression ABSTRACT: Objective To construct recombinant Bb-Em14-3-3 vaccine of Echinococcus multilocularis and to investigate expression efficiency of Em14-3-3 antigen encoding gene in Escherichia coli BL21 (DE3). Methods Em14-3-3 antigen gene was amplified by PCR. Then the gene was cloned into Escherichia coli-Bifidobacteria shuttle plasmid pGEX-1 T containing gluathione-S-transferaze (GST) gene to construct pGEX-Em14-3-3. The recombinant plasmid w