Authors : Salmah Ismail
Abstract: A pure culture of Pasteurella multocida strain B2 has been chosen for shotgun cloning into pUC18 system to determine the virulence genes responsible for haemorrhagic septicaemia (HS) in cow, cattle and buffaloes. Sau3A1 partially digested genomic DNA (2 – 9 kb) from B2 strains was cloned into BamHI site of pUC18. 25 of 790 clones obtained were randomly selected for DNA analysis and mice virulence test. Only one, named as B/392 recombinant plasmid was found to be stable maintained in E. coli host system. The other 23 clones which have tested were found plasmid lost upon subculturing. The plasmid size of B/392 was estimated as 3.6 kb while the size of the inserted fragment was estimated as 0.9 kb (double digested with PstI and SmaI). Clone B/392 showed lethality in mice with at least 100 colony forming unit (CFU) of organisms, and 0.1 ml of the bacterial cultures was injected to cause death within 6 to 24 hours. Further investigation of the B/392 clone was needed to identify the virulence determinant of HS in Pasteurella multocida
Salmah Ismail , 2004. Application of DNA Technology in Determining the Virulence Genes Responsible for Haemorrhagic Septicaemia of Pasteurella multocida Serotype B by Using ‘Shotgun’ Cloning . Asian Journal of Information Technology, 3: 1214-1217.