ABSTRACT

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Cassava (Manihot esculenta) is one of the food crops that contain high carbohydrate and is used as a raw material for various industries. Cassava also contains several types of proteins, including beta-carotene. However, some plants also contain various levels of beta-carotene. Beta-carotene is one of the groups of carotenoids that serve as precursors of vitamin A. This bioinformatics study aims to determine the similarities and differences in the enzyme encoding genes involved in carotenoid biosynthesis pathways in cassava with several plants containing beta-carotene from nucleotide sequences and encoding gene proteins carotenoids (Lcy α and Lcy β). Data on nucleotide and protein sequences were obtained through the NCBI database. Similarities and differences in nucleotide and protein sequence data are seen from the phylogenetic tree using the MEGA 7.0 program. Based on the nucleotide gene Lcy α Manihot esculenta with several plants containing beta-carotene is not related. Based on the phylogenetic tree nucleotide sequence protein Lcy α and Lcy β Manihot esculenta with several plants containing beta-carotene there were differences in similarity

SUMMARY:The development of a dengue virus vaccine is a major priority in efforts to control the diseases. Several researchers are currently using the Asian 1 and Asian 2 genotypes as vaccine candidates for dengue type 2 virus (DENV-2). However, in this study, we constructed a recombinant plasmid-based prM/E gene, from a DENV-2 Cosmopolitan genotype strain as a dengue DNA vaccine candidate. The protein expression of the recombinant plasmid in CHO cells was analyzed using an enzyme-linked immunosorbent assay, western blotting, and sucrose gradient sedimentation. After being used to immunize ddY mice three times at doses of 25 or 100 mg, the DNA vaccine induced humoral immune responses. There was no difference in the neutralizing antibody titer (focus reduction neutralization test 50z value) of mice immunized with 25 and 100 mg DNA vaccine doses. When challenged with 3 × 10 5 FFU DENV-2, immunized mice could raise anamnestic neutralizing antibody responses, which were observed at day 4 and day 8 post-challenge. Analysis of immunogenicity using BALB/c mice showed that their antibody neutralization titers were lower than those of ddY mice. In addition, the antibodies produced after immunization and challenge could also neutralize a DENV-2 Asian 2 genotype (New Guinea C) strain. Therefore, the DENV-2 Cosmopolitan genotype may be a DENV-2 vaccine candidate.