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Abstract
The baculovirus expression vector system (BEVS) has been used as a preferred platform for the production of recombinant protein complexes and efficacious vaccines. However, limited protein yield hinders the application of BEVS. It is well accepted that transcription enhancers are capable of increasing translational efficiency of mRNAs, thereby achieving better protein production. In this study, the ability of LvYY1 as a transcription enhancer was assessed. LvYY1 could interact with the WSSV ie1 promoter via binding to special DNA sites in BEVS. The effects of LvYY1 on protein expression mediated by WSSV ie1 promoter of BEVS was investigated using eGFP as a reporter gene. Enhanced eGFP expression was observed in Sf-9 cells with LvYY1. On this basis, a modified vector combining ie1 promoter and LvYY1 was developed to express either secreting CSFV E2 or baculovirus surface displayed H5 HA of AIVs. Compared to control groups without LvYY1, E2 protein yield increases to 1.6-fold, while H5 production improves as revealed by an upregulated hemagglutination titer of 8-fold at least. Moreover, with LvYY1, H5 displaying baculovirus driven by WSSV ie1 promoter (BV-LvYY1-ie1-HA) sustains the transduction activity in CEF cells. In chicken, BV-LvYY1-ie1-HA elicits a robust immune response against H5 AIVs in the absence of adjuvant, as indicated by specific antibody and cytokine responses. The findings suggest its potential function as both a vectored and subunit vaccine. These results demonstrate that the coexpression with LvYY1 serves as a promising strategy to extensively improve the efficiency of BEVS for efficacious vaccine production.
Highlights
The baculovirus expression vector system (BEVS) has been used as a powerful tool for vaccine development [1,2]
Total RNAs were extracted using an RNAsimple Total RNA Kit (Tiangen, Beijing China) and were reverse transcribed into cDNA using HiScript Q RT SuperMix for Quantitative PCR (qPCR) (Vazyme Biotech, Piscataway, NJ, USA). cDNA was used as the template for qPCR by using the primers (Table 1) targeting the HA5 genomic region
Protein sequences alignment showed that LvYY1 contains four highly conserved zinc fingers and recruit polycomb (REPO) domains like other proteins in the Yin Yang1 (YY1) family (Figure 2A)
Summary
The baculovirus expression vector system (BEVS) has been used as a powerful tool for vaccine development [1,2]. There are other advantages of the baculovirus expression system, including expression of large proteins which are usually correctly folded and biologically active, efficient cleavage of signal peptides, and processing of the protein. BEVS has become the most widely used system in the production of recombinant protein. BEVS allows the production of different vaccine types, including secreting protein and viral surface displayed antigen.
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