Efficient delivery of the immunodominant genes of African swine fever virus by adeno-associated virus serotype 2.

Background and Aim: Adeno-associated virus serotype 2 (AAV2) represents a promising basis for developing a virus-vector vaccine against African swine fever (ASF). This study aimed to create genetic constructs based on AAV2 to deliver the immunodominant genes of ASF virus (ASFV) and to evaluate their functionality in vitro. The efficiency and specificity of transgene expression, as well as their non-toxicity in cells of target animals, were evaluated. Materials and Methods: Bioinformatics analysis methods were used to identify the immunodominant genes of ASFV. The target genes B646L, E183L, CP204L, and CP530R were identified and subsequently cloned into the pAAV-MCS vector. Assembly of recombinant AAV2 (rAAV2) was performed by cotransfection of AAV293 cells with the following plasmids: pAAV-MCS with the gene of interest, envelope, and packaging. Quantitative polymerase chain reaction was used to determine the AAV2 titer. The functionality of the constructs was evaluated in HEK293 and SPEV cells by determining the presence of mature proteins in the cell lysate and the expression levels of messenger RNA. The specificity of the target proteins in cell lysates was confirmed by Western blotting. Results: The proposed AAV2 assembly protocol makes it possible to achieve a concentration of mature viral particles of at least 280 billion/mL of virus-containing material. The rAAV2 could effectively transduce host SPEV cells. The expression of both cistrons was detectable during the transduction of cells; therefore, the combined expression of immunogens in the cells of target animals should be possible using this method. Conclusion: This study demonstrated the potential of using genetic constructs based on AAV2 for the delivery of ASFV genes in vitro.

Effect of concentrated light on morphology and vibrational properties of boron and tantalum mixtures.

Heating a mixture of boron (impurities: carbon ∼ B50C2, boric acid - H3BO3) and tantalum (Ta) powders in nitrogen flow in a xenon high-flux optical furnace was performed. As-received powder composed of h-BN, H3BO3, TaB2, B9H11 and a number of other phases including ß-rhombohedral boron, apparently, heavily doped with Ta. FT-IR examination of any sample of the material reveals the complicated vibration spectrum containing, in particular, an absorption band near 2260 cm-1. The shapes of these bands are different for samples because powders were synthesized at different temperatures. Known, that in ß-rhombohedral boron lattice, there are nano-sized voids of different types, which allow an accommodation of single atoms or small groups of atoms. Theoretical calculations performed by the method of quasi-classical type yields the same value, 2260 cm-1, for the vibrations frequency of Ta atoms in D-type crystallographic voids in ß-rhombohedral boron lattice. Since, Ta atoms are known to prefer accommodation just in D-voids the experimentally detected bands can be identified with localized vibrations of Ta atoms.