Genome Sequence and Phenotypic Analysis of a Protein Lysis-Negative, Attenuated Anthrax Vaccine Strain.

is a Gram-positive bacterium that causes the zoonotic disease anthrax. Here, we studied the characteristic phenotype and virulence attenuation of the putative No. II vaccine strain, PNO2, which was reportedly introduced from the Pasteur Institute in 1934. Characterization of the strain showed that, compared with the control strain, A16Q1, the attenuated PNO2 (PNO2D1) was phospholipase-positive, with impaired protein hydrolysis and significantly reduced sporulation. Additionally, PNO2D1 significantly extended the survival times of anthrax-challenged mice. An evolutionary tree analysis revealed that PNO2D1 was not a Pasteur strain but was more closely related to a Tsiankovskii strain. A database comparison revealed a seven-base insertion mutation in the gene. Although it did not block transcription, the insertion mutation resulted in the premature termination of protein translation. deletion of A16Q1 resulted in a nonproteolytic phenotype that could not sporulate. The database comparison revealed that the gene is also prone to mutation, and the promoter activity was much lower in PNO2D1 than in A16Q1. Low expression may be an important reason for the decreased virulence of PNO2D1.