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| HilD mediated expression of ssrAB based on GFP in Salmonella typhimurium |
| HAN Min-min, ZHANG Li, ZHOU Ni-ni, CHEN Chuan-rong, CAO Kun, LI Yu |
| College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China |
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Abstract Salmonella typhimurium (S.typhi) labeled with GFP was used as the research object and Caenorhabditis elegans as a model organism, initially the role of HilD in the regulation of the expression of ssrAB was explored in vivo. Genetic engineering technology was used to construct S.typhi(ssrAB-gfp+pcDNA3.1), hilD gene deletion strain S.typhi(ΔhilD+ssrAB-gfp+pcDNA3.1) and hilD gene reversion strain S.typhi(ΔhilD+ssrAB-gfp+pcDNA3.1-hilD) carrying GFP gene respectively. Fluorescence microscopy was used to observe the expression and distribution of GFP in 3 labelled strains in Caenorhabditis elegans in vitro and in vivo, and the fluorescence intensity was compared by ImegJ. Three labelled strains were successfully constructed and showed green fluorescence in Caenorhabditis elegans in vitro and in vivo. The fluorescence intensity of S.typhi(ssrAB-gfp+pcDNA3.1) was higher than that of (ΔhilD+ssrAB-gfp+pcDNA3.1-hilD)(P<0.05) and S.typhi(ΔhilD+ssrAB-gfp+pcDNA3.1)(P<0.01), the fluorescence intensity of S.typhi(ΔhilD+ssrAB-gfp+pcDNA3.1-hilD) was higher than that of S.typhi(ΔhilD+ssrAB-gfp+pcDNA3.1)(P<0.05). Colonization of 3 labelled strains in Caenorhabditis elegans in vitro was mainly distributed in oropharyngeal and intestinal tract, the fluorescence intensity of oropharynx was high that of intestinal (P<0.05). The expression of GFP was stable in Salmonella typhimurium, expressing the same in Caenorhabditis elegans. Three labelled strains colonized in oropharyngeal and intestinal very well in Caenorhabditis elegans. The study proved that HilD mediated the expression of ssrAB at the first time in vitro, not only a regulation role.
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Received: 10 January 2018
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| Fund:Supported by the National College Students’ Innovation Training Project (No. 201410364003), the Anhui Natural Science Foundation Project (No. 1508085MC44), and the Provincial Natural Science Research Projects of Anhui Higher Education Institutions (No. KJ2008B056) |
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Corresponding Authors:
Li Yu, Email: liyouer@163.com
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2018, Vol. 34 
