Construction and identification of TSOL18 recombinant Lactococcus lactis secretory and non-secretory vaccines of Taenia solium / 中华地方病学杂志

ABSTRACT

Objective To construct recombinant non-secretory pMG36e-TSOL18/L.lactis and secretory pMG36e-SP-TSOL18/L.lactis vaccines of Taenia solium. Methods Taking sequence of Taenia solium TSOL18 gene as template, genetic optimization was carried out using lactic acid bacteria as a host system, TSOL18 gene and SP-TSOL18 gene were synthesized using PCR-based accurate synthesis (PAS), through the design of full-length primers, the addition of restriction enzyme cutting sites Sac Ⅰ and Hind Ⅲ and protective bases, and the SPUSP45 secretory signal peptide sequence in the N terminal. TSOL18 gene and SP-TSOL18 gene were cloned into Escherichia coli-L.lactis shuttle expression plasmid pMG36e to construct intracellular expression vector pMG36e-TSOL18 and secreted expression vector pMG36e-SP-TSOL18. The two recombinant plasmids were identified by enzyme digestion and sequencing, and electroporated into L.lactis MG1363 to construct the recombinant pMG36e-TSOL18/L.lactis and pMG36e-SP-TSOL18/L.lactis vaccines of Taenia solium, and they were identified by PCR. Results TSOL18 gene fragment and pMG36e vector fragment were obtained by double digestion with Sac Ⅰ and Hind Ⅲ, which were consistent with the expected results; TSOL18 gene standard sequence was aligned and the matching degree was 100％, and both were inserted into Sac Ⅰ and Hind Ⅲ of pMG36e vector. Our PCR results showed that both recombinant pMG36e-TSOL18/L.lactis and pMG36e-SP-TSOL18/L.lactis were 393 bp gene fragment products. Conclusion The recombinant pMG36e-TSOL18/L.lactis and pMG36e-SP-TSOL18/L.lactis vaccines of Taenia solium are successfully constructed.