Cloning And Expression Of The 43ku Gene Of Trichinella From Swine In Hei Longjiang Province

Trichinosis is a kind of serious zoonosis in the world caused by Trichinella spiralis. Ts43 isthe main functionality composition of the ES antigen produced during Trichinella spiralis musclelarvae stage, and it also plays an important part in form of nurse cells. The Ts43 gene was achievedby RT-PCR, The result showed that the cloned gene was closely related with referred sequence with99.61% nucleotide identity and 99.13% derived amino acid identity respectively, furthemoreprokaryotic expression vector pET32-Ts43 and eukaryotic expression vectorpcDNA3.1/CT-GFP-Ts43 were constructed.The prokaryotic expression vector pET32-Ts43 and eukaryotic expression vectorpcDNA3.1/CT-GFP-Ts43 were constructed successfully. The expressed fusion protein by BL-21was pured and renatured, ELISA showed the immunogenicity of the fusion protein was improved alitter bit better after the renaturation. The immunogenicity of renatured fusion protein expressed byBL-21 was not good. The GFP-tagged Ts43 construction was expressed as a recombinant protein inVero E6 cells. Western-blotting showed the recombinant protein was recognized by the mouseantiserum of T. spiralis.The authors consider that Ts43 structural gene is conservative, and the gene can be used toidentify isolates of difficult to identify in morphology. The recombinant protein expressed in VeroE6 cells is a very good candidate for development of T. spiralis recombinant antigen and hasintriguing potential application for the detection and immunization of T. spiralis. It's a means gainedTs43 natural protein too.