Onohuean Hope, Nwodo Uchechukwu U
Biopharmaceutics Unit, Department of Pharmacology and Toxicology, School of Pharmacy, Kampala International University Western Campus, Ishaka-Bushenyi, Uganda.
Biomolecules, Metagenomics, Endocrine and Tropical Disease Research Group (BMETDREG), Kampala International University, Western Campus, Ishaka-Bushenyi, Uganda.
J Genet Eng Biotechnol. 2023 Oct 6;21(1):94. doi: 10.1186/s43141-023-00554-1.
Vibrio species are among the autochthonous bacterial populations found in surface waters and associated with various life-threatening extraintestinal diseases, especially in human populations with underlying illnesses and wound infections. Presently, very diminutive information exists regarding these species' mutational diversity of virulence and resistance genes. This study evaluated variations in endonucleases and mutational diversity of the virulence and resistance genes of Vibrio isolates, harboring virulence-correlated gene (vcgCPI), dihydropteroate synthase type 1 and type II genes (Sul 1 and 11), (aadA) aminoglycoside (3'') (9) adenylyltransferase gene, (aac(3)-IIa, (aacC2)a, aminoglycoside N(3)-acetyltransferase III, and (strA) aminoglycoside 3'-phosphotransferase resistance genes.
Using combinations of molecular biology techniques, bioinformatics tools, and sequence analysis.
Our result revealed various nucleotide variations in virulence determinants of V. vulnificus (vcgCPI) at nucleotide positions (codon) 73-75 (A → G) and 300-302 (N → S). The aminoglycosides resistance gene (aadA) of Vibrio species depicts a nucleotide difference at position 482 (A → G), while the aminoglycosides resistance gene (sul 1 and 11) showed two variable regions of nucleotide polymorphism (102 and 140). The amino acid differences exist with the nucleotide polymorphism at position 140 (A → E). The banding patterns produced by the restriction enzymes HinP1I, MwoI, and StyD4I showed significant variations. Also, the restriction enzyme digestion of protein dihydropteroate synthase type 1 and type II genes (Sul 1 and 11) differed significantly, while enzymes DpnI and Hinf1 indicate no significant differences. The restriction enzyme NlaIV showed no band compared to reference isolates from the GenBank. However, the resistant determinants show significant point nucleotide mutation, which does not produce any amino acid change with diverse polymorphic regions, as revealed in the restriction digest profile.
The described virulence and resistance determinants possess specific polymorphic locus relevant to pathogenomics studies, pharmacogenomic, and control of such water-associated strains.
弧菌属是地表水中原生细菌种群之一,与各种危及生命的肠道外疾病相关,尤其是在有基础疾病的人群和伤口感染中。目前,关于这些物种毒力和耐药基因的突变多样性的信息非常少。本研究评估了携带毒力相关基因(vcgCPI)、二氢蝶酸合酶I型和II型基因(Sul 1和11)、(aadA)氨基糖苷(3'')(9)腺苷酸转移酶基因、(aac(3)-IIa、(aacC2)a、氨基糖苷N(3)-乙酰转移酶III和(strA)氨基糖苷3'-磷酸转移酶耐药基因的弧菌分离株的核酸内切酶变异以及毒力和耐药基因的突变多样性。
使用分子生物学技术、生物信息学工具和序列分析的组合。
我们的结果显示,创伤弧菌(vcgCPI)毒力决定因素在核苷酸位置(密码子)73 - 75(A→G)和300 - 302(N→S)存在各种核苷酸变异。弧菌属的氨基糖苷耐药基因(aadA)在位置482(A→G)处存在核苷酸差异,而氨基糖苷耐药基因(sul 1和11)显示出两个核苷酸多态性可变区域(102和140)。在位置140(A→E)处存在氨基酸差异与核苷酸多态性相关。限制性内切酶HinP1I、MwoI和StyD4I产生的条带模式显示出显著差异。此外,蛋白质二氢蝶酸合酶I型和II型基因(Sul 1和11)的限制性酶切也存在显著差异,而DpnI和Hinf1酶显示无显著差异。与来自GenBank的参考分离株相比,限制性内切酶NlaIV未显示条带。然而,如限制性酶切图谱所示,耐药决定因素显示出显著的点核苷酸突变,在不同的多态性区域未产生任何氨基酸变化。
所描述的毒力和耐药决定因素具有与病原组学研究、药物基因组学以及此类与水相关菌株的控制相关的特定多态性位点。