Gee P, Maron D M, Ames B N
Division of Biochemistry and Molecular Biology, University of California, Berkeley 94720-3202.
Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11606-10. doi: 10.1073/pnas.91.24.11606.
A detection and classification system for mutagens has been developed that identifies the six possible base-pair substitution mutations. A set of six Salmonella typhimurium (TA7001 to TA7006) strains has been constructed, each of which carries a unique missense mutation in the histidine biosynthetic operon. In addition to the his mutation, these strains carry different auxiliary features that enhance the mutability of the target his mutation. These include the R factor pKM101, which has the SOS-inducible mucAB system; a deletion of the uvrB component of excision repair; and rfa mutations to increase the accessibility of bulky chemicals to the bacteria. Another set of strains (TA7041 to TA7046) contain a wild-type rfa gene. Reversion via the base substitution unique to each strain was verified by sequence analyses of > 800 revertants obtained from different types of mutagens. The strains have considerably lower spontaneous reversion frequencies and detect a variety of mutagens at a sensitivity comparable to the Salmonella tester strains TA100, TA102, and TA104. The low spontaneous frequency of reversion of a mixture of the six tester strains (approximately 10 revertants per plate) enables a single mutation assay with the mixture that is followed by classification of the type of mutation with the individual strains.
已开发出一种用于诱变剂的检测和分类系统,该系统可识别六种可能的碱基对替换突变。构建了一组六种鼠伤寒沙门氏菌(TA7001至TA7006)菌株,每种菌株在组氨酸生物合成操纵子中携带独特的错义突变。除了his突变外,这些菌株还具有不同的辅助特征,可增强目标his突变的易变性。这些特征包括具有SOS诱导型mucAB系统的R因子pKM101;切除修复的uvrB成分缺失;以及rfa突变,以增加大分子化学物质对细菌的可及性。另一组菌株(TA7041至TA7046)含有野生型rfa基因。通过对从不同类型诱变剂获得的800多个回复突变体进行序列分析,验证了每种菌株特有的碱基替换导致的回复突变。这些菌株的自发回复频率相当低,并且以与沙门氏菌测试菌株TA100、TA102和TA104相当的灵敏度检测多种诱变剂。六种测试菌株混合物的低自发回复频率(每平板约10个回复突变体)使得能够对该混合物进行单一突变检测,随后用单个菌株对突变类型进行分类。