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vc小小子铁杆木虫 (著名写手)
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[求助]
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摘要： 为了研究鼠伤寒沙门菌crp缺失株对HeLa细胞黏附和侵入的影响，本试验对培养好的鼠伤寒沙门菌SL1344和SL1344Δcrp株进行连续稀释，随后涂板计数，计算原菌液的CFU；对培养好的HeLa细胞调至每孔4×105 CFU，然后接种于6孔板中培养。将处理好的细菌悬液以感染复数（MOI）100:1加入到6孔板中，培养2小时，PBS洗涤后用胰酶消化，然后取细胞悬液100 μL连续10倍稀释，挑选3个稀释度进行涂板计数，计算黏附率；.而侵入试验是在加入6孔板培养2 h后，用含10%血清和100 μg/mL的庆大霉素的DMEM培养基培养培养1.5 h，然后用裂解液处理该细胞，接着进行对比稀释，挑选3个梯度进行涂板计数，计算侵入率。试验结果显示，在黏附试验中，SL1344菌株的黏附率为0.155%，SL1344Δcrp株的黏附率为0.12%，差异显著（P<0.05）。在侵入试验中，SL1344菌株的侵入率为0.1%，而SL1344Δcrp的侵入率为0.08%，两者的侵入率相比差异显著（P<0.05）。结果表明，crp缺失株对HeLa细胞的黏附率与侵袭率均发生降低，表明crp缺失株的致病力发生了减弱，本试验为研制安全有效的鼠伤寒沙门菌疫苗以及疫苗载体奠定了基础。 关键词：鼠伤寒沙门菌，crp，黏附，侵入 |
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灿灿妹
至尊木虫 (著名写手)
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| The cultured HeLa cells were adjusted to 4×105 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 μL 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 μg/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. |
3楼2015-05-31 09:22:39
灿灿妹
至尊木虫 (著名写手)
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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 SL1344Δcrp) of Salmonella typhimurium were continuous diluted, followed by coated plates to calculate CFU of the original bacterium. |
2楼2015-05-31 08:50:51
灿灿妹
至尊木虫 (著名写手)
- 翻译EPI: 68
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- 性别: GG
- 专业: 环境工程
【答案】应助回帖
★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★
vc小小子: 金币+35, ★★★很有帮助 2015-06-02 11:22:33
vc小小子: 金币+35, ★★★很有帮助 2015-06-02 11:22:33
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Experimental results showed that, for adhesion test, the adhesion rate of SL1344 and SL1344Δcrp 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 |
4楼2015-05-31 09:38:54
