Construction and identification of the IL-35-BCG recombinant vaccine

ObjectiveTo construct the recombinant eukaryotic expression vector pZM03 (pBudCE4.1/GLS/IL35/OriM) containing secretory protein of interleukin 35 (IL-35)-bacillus calmette guerin (BCG)recombinant vaccine.

MethodsThe chemically synthesized IL-35 gene fragment was cloned into the pBudCE4.1/GLS/OriM, the pBudCE4.1/GLS/IL35/OriM was obtained from the pBudCE4.1/GLS/IL12/OriM, and both of above was used to construct the co-expression vector pZM03 (pBudCE4.1/GLS/IL35/OriM).The recombinant plasmid pZM03 was electrotransformated into the BCG to construct the recombinant co-expression of IL-35 and BCG plasmid pBudCE4.1/GLS/IL35/OriM-BCG.

ResultsThe results of the detection of gene, transcription, translation and cellular levels, which included the PCR, sequencing, Western blotting and acid-fast staining methods, showed that the pBudCE4.1/GLS/IL35/OriM-BCG was successfully constructed.

ConclusionThe recombinant plasmid pBudCE4.1/GLS/IL35/OriM-BCG can correctly express IL-35, which lays a good foundation for further research on the function and application value of IL-35.