The porcine reproductive and respiratory syndrome (PRRS) caused the serious economic damage to swine breeding around the world. It is a viral infective disease against which live attenuated and inactivated vaccines are not always successful. Development of new types of drugs such as DNA vaccines is necessary for improving the protection against the virus. DNA vaccines induce the development of both a cellular and humoral immune response. Such vaccines consist of a plasmid or viral vector with genes of potentially immunogenic proteins. The expression of these genes realized in cells of the vaccinated animal. It leads to the synthesis of antigen proteins triggering the immune response. The purpose of this work is to create a genetic construction that can be used as DNA vaccine against PRRS virus. The construction consists of the commercial vector pVAX1 and open reading frame of two structural proteins of PRRS virus, a lysosomal localization signal sequence of the invariant chain gene and regulatory elements necessary for the expression of cloned genes in mammalian cells.
To improve expression of the porcine circovirus type 2 (PCV2) capsid protein in E. coli cells, the corresponding gene was optimized and two variants of the open reading frame were constructed, which encoded the full-sized and shortened capsid proteins as part of the expression vector. Rare codons were replaced, and in the case of a shortened version of the gene, the region corresponding to the N-terminal domain of the protein was deleted. A comparison was made of the expression level of the studied proteins. It was established that the highest level of expression in bacterial cells is achieved by simultaneously optimizing the codons and removing the initial (N-terminal) 108 base pair (bp) portion of the gene, which contains the nuclear localization signal.
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