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. 2017 Jul 15;25(10):2332–2344. doi: 10.1016/j.ymthe.2017.06.017

Figure 2.

Figure 2

Construction and In Vitro Characterization of SCV-CHIK

(A) Schematic representation of SCV-CHIK construction. The CHIKV structural polyprotein cassette (C-E3-E2-6K-E1) was inserted into the A39R locus of VACV to generate VACV-CHIK. The D13L gene was deleted from VACV-CHIK to generate SCV-CHIK. (B) Viral DNA from parental VACV and SCV-CHIK was subjected to PCR analysis using site-specific primers shown in (A) to confirm the insertion of C-E3-E2-6K-E1 (green arrow) and deletion of D13L (blue arrow). (C) Immunoblot analysis of VACV- and SCV-CHIK-infected SCS cells using mouse polyclonal anti-CHIKV antisera confirmed expression of processed CHIKV antigens: pE2 (E3-E2), E1 and E2 (which co-migrate), and capsid.