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目的检测重组人Β防御素2(HUMANΒ-DEFENSIN2,HBD2)腺病毒表达载体在大鼠真皮多能干细胞(DERMALMULTIPOTENT STEM CELLS,DMSCS)中的表达,并观察重组HBD2的体外抗菌活性。方法将含有HBD2重组腺病毒转染DMSCS,RT-PCR、荧光免疫化学、WESTERN BLOTTING检测HBD2的表达情况,ELISA测定培养上清中HBD2的浓度,K-B纸片扩散法检测上清对大肠埃希菌、金黄色葡萄球菌和铜绿假单胞菌等标准菌株的杀灭效果。结果RT-PCR、荧光免疫化学和WESTERNBLOTTING的结果显示转染后HBD2可在DMSCS中有效地表达,上清中HBD2的浓度为743.6NG/ML,K-B纸片法显示HBD2对上述标准菌株有明显的杀灭效应。结论HBD2重组腺病毒表达载体在DMSCS可高效表达,并对大肠埃希菌等标准菌株有杀灭效应。
Objective To detect the expression of recombinant human β-defensin 2 (HBD2) adenovirus in DERMALMULTIPOTENT STEM CELLS (DMSCS) and to observe the antibacterial activity of recombinant HBD2 in vitro. Methods The expression of HBD2 was detected by RT-PCR, immunofluorescence and WESTERN BLOTTING. The HBD2 concentration in the culture supernatant was determined by ELISA. The supernatant was tested for the effect of Escherichia coli , Staphylococcus aureus and Pseudomonas aeruginosa and other standard strains of killing effect. Results The results of RT-PCR, immunofluorescence and WESTERNBLOTTING showed that HBD2 could be efficiently expressed in DMSCS after transfection. The concentration of HBD2 in the supernatant was 743.6NG / ML. The KB disk method showed that HBD2 had obvious Kill effect. Conclusion HBD2 recombinant adenovirus expression vector in DMSCS can be highly expressed, and Escherichia coli and other standard strains kill effect.