1887

Abstract

is a prominent human pathogen and is known to form L-forms and during infection. However, the conditions of L-form formation are not optimal and the mechanisms of L-form formation in this organism are unknown. Here, we optimized the conditions of unstable L-form formation, constructed a transposon mutant library and screened for mutants defective in unstable L-form formation. Our results revealed that 20 % sucrose, 3.5 % sodium chloride, 750–1000 U penicillin and 33 °C were optimal conditions for L-form formation. Stationary phase cultures of formed L-forms better than exponential phase cultures. The L-form colonies showed typical ‘fried-egg’ morphology and the cells had deficient cell wall, showed morphological diversity, and stained Gram-negative. The mutant library screens identified 15 mutants deficient in L-form formation and sequencing analysis identified mutations in eight genes and three intergenic regions. Real-time PCR analysis indicated that, with the exception of , seven genes including , , , , , and were preferentially expressed in L-forms as compared with normal cell-walled form (<0.05). The identified genes involved in L-form growth mapped in the pathways for energy production, iron homeostasis, transporters, DNA repair, membrane biogenesis, and biosynthesis. Our findings shed new insight into the molecular basis of unstable L-form formation and may have implications for development of novel drugs targeting L-forms for improved treatment.

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2015-01-01
2021-07-29
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