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For investigating the effects of Salmonella typhimurium crp nullmutant to the adhesion and invasion of HeLa cell,two cultured strains (namely  SL1344 and SL1344¦¤crp) of Salmonella typhimurium were continuous diluted, followed by coated plates to calculate CFU of the original bacterium.
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The cultured HeLa cells were adjusted to 4¡Á105 CFU per hole, then were  inoculated into 6-well plate, where the treated bacterial suspension was  added into soon afterwards , as MOI was 100:1. After 2h cultivation, the 6-well plate was washed by PBS and digested by  trypsin to fetch 100 ¦ÌL bacterial suspension, which was continuous diluted 10 times, and 3 dilution were selected to coated plate count to calculate adhesion rate. While for invasion test, the 6-well plate were cultivated again by DMEM culture medium, whcih was composed by 10% serum and 100 ¦Ìg/mL gentamicin, after 2h cultivation. And then the cell was treated by lysate, followed by compared dilution to select  3 dilution,which were used for coated plate counting to calculate invasion efficiency.
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3Â¥2015-05-31 09:22:39
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vcСС×Ó: ½ð±Ò+35, ¡ï¡ï¡ïºÜÓаïÖú 2015-06-02 11:22:33
Experimental results showed that, for adhesion test, the adhesion rate of SL1344 and SL1344¦¤crp were 0.155% and 0.12% respectively, with significant difference £¨P<0.05£©. While for invasion experiment, its  invasion efficiency wer 0.1% and 0.08% separately, with significant different £¨P<0.05£©.
The results showed that  adhesion rate and invasion efficiency of crp nullmutant to HeLa cell were degradation, which indicated that its  virulence has weaken. This experiment has established the base of developing safe and effective Salmonella typhimurium vaccine and vaccine vector.

Key words:  Salmonella typhimurium, crp,  adhesion,  invasion
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