培养基中氯化钠对百日咳杆菌CS株生长及百日咳毒素表达的影响  

Effects of sodium chloride in medium on growth of Bordetella pertussis and expression of pertussis toxin

作　　者：李应伟[1] 刘晓凤[1] 周晓舟[1] 夏永[1] 

机构地区：[1]成都生物制品研究所有限责任公司,四川成都610023

出　　处：《中国生物制品学杂志》2015年第11期1138-1141,共4页Chinese Journal of Biologicals

基　　金：国家"十二五"重大新药创制专项(2013ZX09402-302-005)

摘　　要：目的探讨培养基中氯化钠（Na Cl）对百日咳杆菌CS株生长及百日咳毒素（pertussis toxin,PT）表达的影响,确定最适PT表达的Na Cl加入方式。方法以MSS为起始培养基,补加不同浓度的Na Cl溶液,接种生产用百日咳杆菌Ⅰ相CMCC58003（CS株）后,分别于不同时间取样;分别于接种菌种后不同时间补加Na Cl至7.5 g/L,于培养结束时（39 h）取样;分别于接种菌种后0和25 h时补加Na Cl至7.5 g/L,培养不同时间取样。上述样品分别测定菌浓度及PT表达量。结果起始培养基中Na Cl浓度越高,细菌生长越缓慢,菌浓度增加时间也越长,起始培养基中Na Cl浓度为7.5 g/L时,培养上清中PT表达量最高,达6.61μg/ml,比对照培养基（MSS）提高了15.6%。培养过程中在25 h时补加Na Cl,培养结束时PT表达量最高,为8.64μg/ml,比对照培养基（MSS）提高了43.28%,比0 h时补加Na Cl提高了26.13%。在25 h时补加Na Cl后,细菌的菌浓度增长期比MSS培养基培养延长了2~3 h,比0 h时补加Na Cl缩短了1~2 h,菌浓度在培养的中后期明显高于0 h时补加Na Cl,PT表达周期与生长曲线基本平行,当细菌生长进入衰退期时,PT也随之开始逐步降解。结论在培养前期,可采用含2.5 g/L Na Cl的MSS培养基使菌体快速生长,当进入抗原PT表达期后,可通过补加Na Cl来提高PT转录活性,从而提高PT的产量。Objective To investigate the effect of sodium chloride in medium on the growth of Bordetella pertussis and the expression of pertussis toxin（PT）so as to optimize the mode of addition of sodium chloride which was more beneficial to PT expression.Methods MMS used as an initial medium was added with sodium chloride solution at various concentrations,and inoculated with B.pertussis phaseⅠ CMCC58003（CS strain）for production,from which samples were taken at various time points.The medium was supplemented with sodium chloride to a concentration of 7.5 g / L at various time points after inoculation,from which samples were taken at end of culture（39 h）.Meanwhile,the medium was supplemented with sodium chloride to a concentration of 7.5 g / L 0 and 25 h after inoculation respectively,from which samples were taken at various time points.All the samples were determined for bacterial concentration and PT expression level.Results The higher the sodium chloride concentration in initial medium was,the slower bacteria grew,and the longer the increasing time of bacterial concentration was.When the sodium chloride concentration in initial medium was7.5 g / L,the PT expression level in culture supernatant reached a peak value of 6.61 μg / ml,which increased by15.6% as compared with that in control medium（MSS）.However,the PT expression level at end of culture reached a peak value of 8.64 μg / ml when the sodium chloride was supplemented 25 h after inoculation,which increased by43.28% as compared with that in control medium（MSS）,and by 26.13% as compared with that in the medium supplemented with sodium chloride 0 h after inoculation.The increasing time of bacterial concentration in the medium supplemented with sodium 25 h after inoculation was 2 ~ 3 h longer than that in control medium MSS,and 1 ~ 2 h shorter than that in the medium supplemented with sodium chloride 0 h after inoculation.The bacterial concentrations at middle and late stages of culture were significantly higher than that in the medium supplemen

关 键 词：MSS培养基 氯化钠 百日咳杆菌 百日咳毒素 

分 类 号：R378.42[医药卫生—病原生物学] R392-33[医药卫生—基础医学]