DNA amplification optimization and cloning of several target genes from burkholderia pseudomallei

Abdul Latif Osman, Syed Mohamed Syed Abdullah, Siti Marhamah Drahaman, Mohd Firdaus Mohd Raih, Noraslinda Muhamad Bunnori, Aisyah Mohamed Rehana

Research output: Contribution to journalArticle

Abstract

Burkholderia pseudomallei is a saprophytic Gram-negative bacillus that is found in soil and surface water. It causes the disease melioidosis, which infect humans and animals. Melioidosis can be fatal in human, where it causes fever and commonly present with pneumonia, with or without septicaemia. Melioidosis is primarily endemic in Southeast Asia and Northern Australia, but has also been found in the Middle East, China and South America. It is challenging to treat melioidosis, as it is intrinsically resistant to many antibiotics, and can cause latent infection. By characterizing identified protein targets from B. pseudomallei, we can gain fundamental knowledge on how this bacterium behaves, and thus provide us strategies to combat them. We report here the recent progress of DNA amplification and cloning of four target genes from B. pseudomallei strain D286 performed in Kulliyyah of Science, IIUM. Genomic DNA of B. pseudomallei strain D286 is obtained from School of Biosciences & Biotechnology, UKM. The four target genes; BPSL1612, BPSL1618, BPSL1691 and BPSL2054 were chosen from 52 hypothetical proteins predicted to be essential in B. pseudomallei by transposon-directed insertion site sequencing (TraDIS) technique (Moule et al., 2014). All four target genes were subjected to nested PCR amplification for subsequent Gateway™ cloning protocols, expression and purification studies.

Original languageEnglish
Pages (from-to)153-158
Number of pages6
JournalJurnal Teknologi
Volume77
Issue number25
DOIs
Publication statusPublished - 2015

Fingerprint

Cloning
Amplification
DNA
Genes
Proteins
Antibiotics
Bacilli
Biotechnology
Surface waters
Purification
Bacteria
Animals
Soils

Keywords

  • B. pseudomallei
  • Gateway™ cloning
  • hypothetical proteins
  • Melioidosis

ASJC Scopus subject areas

  • Engineering(all)

Cite this

Osman, A. L., Abdullah, S. M. S., Drahaman, S. M., Mohd Raih, M. F., Bunnori, N. M., & Rehana, A. M. (2015). DNA amplification optimization and cloning of several target genes from burkholderia pseudomallei. Jurnal Teknologi, 77(25), 153-158. https://doi.org/10.11113/jt.v77.6756

DNA amplification optimization and cloning of several target genes from burkholderia pseudomallei. / Osman, Abdul Latif; Abdullah, Syed Mohamed Syed; Drahaman, Siti Marhamah; Mohd Raih, Mohd Firdaus; Bunnori, Noraslinda Muhamad; Rehana, Aisyah Mohamed.

In: Jurnal Teknologi, Vol. 77, No. 25, 2015, p. 153-158.

Research output: Contribution to journalArticle

Osman, Abdul Latif ; Abdullah, Syed Mohamed Syed ; Drahaman, Siti Marhamah ; Mohd Raih, Mohd Firdaus ; Bunnori, Noraslinda Muhamad ; Rehana, Aisyah Mohamed. / DNA amplification optimization and cloning of several target genes from burkholderia pseudomallei. In: Jurnal Teknologi. 2015 ; Vol. 77, No. 25. pp. 153-158.
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