Construction and investigation of plasmid-induced effects on growth of GFP-expressing Salmonella strains

Ubaidah Naim Taraq Naem Zia, Deepah Samynathan, Hasni Idayu Saidi, G. T. Selvarajah, Y. Cheah
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Abstract

Background: Bacteria such as the Salmonella species had been extensively studied not only for its mechanism of pathogenicity, but for its beneficial utilisation in bacterial-mediated tumour therapy. Genetically modified bacterial strain, BDLA Salmonella Agona (BDLA S. Agona) had shown great promise as tumour targeting and suppressing agent. By, constructing green fluorescent protein (GFP) expressing of this strain, it would be beneficial as it could be used to further study the interaction of the BDLA S. Agona strain with various cancerous cells and the host at the systemic level. However, GFP plasmid-induced strains may exhibit growth defects, and this was investigated in this study. Methods: The GFP-transformed Salmonella strains were constructed, and the replication rate of the strains were investigated using the BacterioScan™ 216R instrument. The replication rate between the strains was compared from the 24-hour data collected and analysed as growth curves. The 24-hour growth curves were constructed using the Log CFU/mL and OD650 data collected by the instrument. Results: It was observed from the growth curves that the transformation of the GFP plasmid into different Salmonella strains did not affect the replication rate of the bacteria. Discussion and Conclusion: This finding answers the objective of the study, and it was concluded that incorporation of the GFP plasmid does not cause any negative effect on bacterial growth. GFP-transformed Salmonella, specifically the BDLA S. Agona strain, could be utilised as a powerful tool for future studies on the mechanism of tumour suppression and real-time in vivo biodistribution of the strain.
质粒对表达gfp的沙门氏菌生长影响的构建及研究
背景:沙门氏菌等细菌不仅因其致病性机制,而且因其在细菌介导的肿瘤治疗中的有益利用而被广泛研究。转基因BDLA沙门氏菌Agona (BDLA S. Agona)作为肿瘤靶向和抑制药物已显示出巨大的潜力。构建表达该菌株的绿色荧光蛋白(GFP),有助于在系统水平上进一步研究BDLA S. Agona菌株与多种癌细胞及宿主的相互作用。然而,GFP质粒诱导的菌株可能表现出生长缺陷,本研究对此进行了研究。方法:构建gfp转化的沙门菌菌株,利用BacterioScan™216R仪器检测菌株的复制率。通过收集的24小时数据比较菌株之间的复制率,并以生长曲线分析。利用仪器采集的Log CFU/mL和OD650数据构建24小时生长曲线。结果:从生长曲线上观察到,GFP质粒在不同沙门氏菌中的转化对细菌的复制率没有影响。讨论与结论:这一发现回答了本研究的目的,并得出结论,GFP质粒的掺入不会对细菌生长产生任何负面影响。gfp转化的沙门氏菌,特别是BDLA S. Agona菌株,可以作为未来研究该菌株肿瘤抑制机制和实时体内生物分布的有力工具。
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