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目的探讨真核表达质粒pIRES2-EGFP-4-1BBL转入对减毒鼠伤寒沙门菌SL3261生物学行为的影响。方法将真核表达质粒pIRES2-EGFP-4-1BBL通过两步电转化获得含pIRES2-EGFP质粒的SL3261,接种于LB培养基中观察其生长特性、革兰染色观察其形态和血清凝集试验检测其表面抗原变化。利用该重组菌体外感染HepG2细胞,观察其对细胞的侵袭能力的影响。结果含pIRES2-EGFP-4-1BBL质粒SL3261在体外生长繁殖较原细菌慢,革兰染色呈阴性丝状菌体,含pIRES2-EGFP-4-1BBL质粒的SL3261 A-F多价、O4菌体和H1鞭毛抗原和SL3261完全一致;体外感染HepG2能力,SL3261为201±46 CFU/200HepG2细胞,SL3261-pIRES2-EGFP为163±37,而SL3261-pIRES2-EGFP-4-1BBL为158±32,3组间的差异无统计意义(P>0.05)。结论导入pIRES2-EGFP-4-1BBL质粒对SL3261的生长和形态有部分影响,而表面抗原以及侵入HepG2细胞的功能不变,该结果为含pIRES2-EGFP-4-1BBL质粒SL3261疫苗菌的进一步研究奠定了实验依据。
Objective To investigate the effect of transfection of eukaryotic expression plasmid pIRES2-EGFP-4-1BBL on the biological behavior of attenuated Salmonella typhimurium SL3261. Methods The eukaryotic expression plasmid pIRES2-EGFP-4-1BBL was transformed into pIRES2-EGFP plasmid SL3261 by electroporation in two steps and inoculated into LB medium to observe its growth characteristics. Gram stain was used to observe its morphology and serum agglutination test Surface antigen changes. HepG2 cells were infected with the recombinant bacteria in vitro to observe their influence on cell invasiveness. Results Plasmid SL3261 containing pIRES2-EGFP-4-1BBL grew more slowly than the original bacteria in vitro with negative stains of Gram stain, SL3261 AF multivalent with pIRES2-EGFP-4-1BBL plasmid, O4 cells and H1 The flagellar antigen was completely identical to SL3261; the ability of HepG2 to infect in vitro was 201 ± 46 CFU / 200HepG2 for SL3261, 163 ± 37 for SL3261-pIRES2-EGFP and 158 ± 32 for SL3261-pIRES2-EGFP-4-1BBL The difference was not statistically significant (P> 0.05). Conclusion The introduction of plasmid pIRES2-EGFP-4-1BBL has some effects on the growth and morphology of SL3261, while the surface antigen and the function of invading HepG2 cells remain unchanged. The result is a further study of SL3261 vaccine containing pIRES2-EGFP-4-1BBL plasmid Laid the experimental basis.