Polymerase chain reaction and cloning of Burkholderia pseudomallei putative genes

Nagi A. Al-Haj, Lai L. Suang, Mariana N. Shamsudin, Rasedee Abdullah, Rahman Mohamed, Zamberi Sekawi

Research output: Contribution to journalArticle

Abstract

Total of 23 putative Open Read Farms from B. pseudomallei strain D286 was successfully cloned and the nucleotide sequence analysis of the putative genes showed the homologue (98-100%) to strain K96243. The high similarity in gene sequences between these strains is confirmed for presence of the necessary ORF for LPS biosynthesis through PCR amplification the application of the ORFs in the PCR amplification and expression method. The findings of this study have contributed to some information on the molecular bases of the LPS biosynthesis genes in B. pseudomallei specifically for strain D286. PCR amplification, a specific pair of primer for each ORFs was proving specific for amplification of genes in B. pseudomallei strain D286. The PCR mixture with addition of DMSO, formamide and glycerol could ease the PCR optimization where different pairs of primers were involved. The specific primer pairs with the PCR mixture could be used in developing a PCR diagnosis of melioidosis.

Original languageEnglish
Pages (from-to)104-110
Number of pages7
JournalResearch Journal of Medical Sciences
Volume3
Issue number3
Publication statusPublished - 2009
Externally publishedYes

Fingerprint

Burkholderia pseudomallei
Organism Cloning
Polymerase Chain Reaction
Genes
Open Reading Frames
Melioidosis
Gene Amplification
Dimethyl Sulfoxide
Glycerol
Sequence Analysis

Keywords

  • Burkholderia pseudomallei
  • Cloning
  • Melioidosis
  • Open reading frames
  • Polymerase chain reactor
  • Primers
  • Putative genes

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Al-Haj, N. A., Suang, L. L., Shamsudin, M. N., Abdullah, R., Mohamed, R., & Sekawi, Z. (2009). Polymerase chain reaction and cloning of Burkholderia pseudomallei putative genes. Research Journal of Medical Sciences, 3(3), 104-110.

Polymerase chain reaction and cloning of Burkholderia pseudomallei putative genes. / Al-Haj, Nagi A.; Suang, Lai L.; Shamsudin, Mariana N.; Abdullah, Rasedee; Mohamed, Rahman; Sekawi, Zamberi.

In: Research Journal of Medical Sciences, Vol. 3, No. 3, 2009, p. 104-110.

Research output: Contribution to journalArticle

Al-Haj, NA, Suang, LL, Shamsudin, MN, Abdullah, R, Mohamed, R & Sekawi, Z 2009, 'Polymerase chain reaction and cloning of Burkholderia pseudomallei putative genes', Research Journal of Medical Sciences, vol. 3, no. 3, pp. 104-110.
Al-Haj NA, Suang LL, Shamsudin MN, Abdullah R, Mohamed R, Sekawi Z. Polymerase chain reaction and cloning of Burkholderia pseudomallei putative genes. Research Journal of Medical Sciences. 2009;3(3):104-110.
Al-Haj, Nagi A. ; Suang, Lai L. ; Shamsudin, Mariana N. ; Abdullah, Rasedee ; Mohamed, Rahman ; Sekawi, Zamberi. / Polymerase chain reaction and cloning of Burkholderia pseudomallei putative genes. In: Research Journal of Medical Sciences. 2009 ; Vol. 3, No. 3. pp. 104-110.
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