Construction of yeast expression vector of rubella virus glycoprotein E1 gene

Objective: To construct the cloning and yeast expressing vectors of rubella virus(RV) glycoprotein E1. Methods: The whole sequence of E1 gene was amplified by PCR, and corresponding restriction enzymes(RE) sites, EcoRⅠ, XbaⅠ, were created in the both ends of the sequence. The gene of E1 and pBluscriptⅡSK+ were digested by EcoRⅠ and XbaⅠ respectively, ligated and transformed into E.coli JM109. After being screened by ampicillin resistant and LacZ+, the products were confirmed by PCR, enzyme digestion and sequence analysis. The cloning vector and the expressing plasmid pGAPZαA-E1 were digested by EcoRⅠ and XbaⅠ, ligated and transformed into E.coli JM109. The products were screened by Zeocin, and after sequence analysis, the pGAPZαA-E1 was constructed. Results: The result of PCR was a fragment of 1.5 kb. The PCR and enzyme digestion results of both recombinant plasmid pBluscriptⅡSK+-E1 and pGAPZαA-E1 showed the correct fragments, and the sequence analysis confirmed that the nucleotide sequence was E1 gene. Conclusion: The recombinant yeast expression vector of RV-E1 was constructed successfully.