Ames试验菌株传代次数对菌株活性和自发回复突变的影响

张慧君; 陶功华; 肖萍

doi:10.19428/j.cnki.sjpm.2020.20278

Ames试验菌株传代次数对菌株活性和自发回复突变的影响

Effect of passage number on the activity and spontaneous reverse mutation of Ames test strains

摘要

摘要:
目的对Ames试验用菌株生物学特性的稳定性进行研究，为确定菌株生物学特性不发生改变的传代次数提供依据。
方法选择Ames试验所需使用的鼠伤寒沙门菌TA97a、TA98、TA100、TA102菌株作为实验菌株。分别使用原代和不同传代次数的冻存菌株进行生物学特性和自发回复突变菌落数的检测。
结果4种菌株F1~F6代的组氨酸缺陷、脂多糖屏障缺损、氨苄青霉素抗性、紫外线敏感性和四环素抗性的生物学特性与F0代相比均无改变。但TA97a菌株的自发回变数在F3代开始与F0的差异有统计学意义(P < 0.05)，而TA100、TA102菌株的自发回变数在F4代开始与F0代的差异有统计学意义(P < 0.05)。
结论Ames试验菌株的传代次数会影响菌株的自发回复突变，TA98、TA100、TA102菌株的传代次数不应超过4代，TA97a菌株的传代次数不应超过3代。

Abstract:
ObjectiveWe explored the stability of the bacteria strains used in the Ames test to provide a basis for determining the appropriate passage number at which the biological characteristics of the strains would not change.
MethodsThe Salmonella typhimurium (TA97a, TA98, TA100 and TA102 strains) were selected as the experimental strains.The original frozen strains and frozen strains with different passage times were used to compare the biological characteristics and the spontaneously reverting colonies.
ResultsThe biological characteristics of four kinds of strains, which were histidine deficiency, lipidpolysaccharide barrier defect, ampicillin resistance, UV sensitivity, and tetracycline resistance, did not change at F1-F6 generation when compared with the F0 generation.However, as for the number of spontaneously reverting colonies, a statistically significant difference (P < 0.05) occurred at F3 generation when compared with F0 generation for the TA97a strain, and a significant difference (P < 0.05) occurred at F4 generation for TA100 and TA102 strains.
ConclusionPassage number of strains used in Ames test could affect their spontaneous reversion mutation rate.The passage number should be less than 4 for TA98、TA100、TA102 strains, and less than 3 for TA97a in Ames test.

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