Martin S M, Shoham S C, Alsup M, Rogolsky M
Infect Immun. 1980 Feb;27(2):532-41. doi: 10.1128/iai.27.2.532-541.1980.
Two regimens for transformation have been devised for the phage group 2 strains UT0002-19 and UT0017 of Staphylococcus aureus. Strain UT0002-19 produces exfoliative toxin, which is responsible for the clinical manifestations of the staphylococcal scalded skin syndrome, whereas strain UT0017 does not produce exfoliative toxin. A large pool of antibiotic-resistant and auxotrophic mutants from strains UT0002-19 and UT0017 were isolated by using (i) antibiotic gradient plates, (ii) trimethoprim selection, and (iii) nitrosoguanidine mutagenesis, which sometimes was coupled by enrichment with penicillin or methicillin. Transformation frequencies of genetic markers in mutant strains ranged from 10(-6) to 10(-8). Three genetic linkage groups were identified on the strain UT0017 chromosome. The first linkage group was thy-4-lys-5-trp-21-thr-4, the second was pyr-26-nov-9-his-3, and the third consisted of ilv-9 and pen-1, a genetic determinant for beta-lactamase synthesis. Two linkage groups were detected on the strain UT0002-19 CHROMOSOMe. The first was defined as thy-1-lys-5-trp-3-thr-4-ala-8, whereas the second consisted of nov-9 and his-3 markers. A chromosomal locus that controlled synthesis of exfoliative toxin could not be mapped.
已为金黄色葡萄球菌的噬菌体2组菌株UT0002 - 19和UT0017设计了两种转化方案。菌株UT0002 - 19产生剥脱毒素,该毒素是葡萄球菌烫伤样皮肤综合征临床表现的病因,而菌株UT0017不产生剥脱毒素。通过使用(i)抗生素梯度平板、(ii)甲氧苄啶筛选和(iii)亚硝基胍诱变从菌株UT0002 - 19和UT0017中分离出大量抗生素抗性和营养缺陷型突变体,亚硝基胍诱变有时会通过用青霉素或甲氧西林富集来进行。突变菌株中遗传标记的转化频率范围为10^(-6)至10^(-8)。在菌株UT0017染色体上鉴定出三个遗传连锁群。第一个连锁群是thy - 4 - lys - 5 - trp - 21 - thr - 4,第二个是pyr - 26 - nov - 9 - his - 3,第三个由ilv - 9和pen - 1组成,pen - 1是β - 内酰胺酶合成的遗传决定因素。在菌株UT0002 - 19染色体上检测到两个连锁群。第一个被定义为thy - 1 - lys - 5 - trp - 3 - thr - 4 - ala - 8,而第二个由nov - 9和his - 3标记组成。控制剥脱毒素合成的染色体位点无法定位。