Penyaringan domain ADP-ribosilasi gen toksin daripada Burkholderia pseudomallei virulen

Translated title of the contribution: Screening ADP-ribosilation domain of toxin gene from Burkholderia pseudomallei virulent

Shahrul Hisham Zainal Ariffin, Rahmah Mohamed, Zulkeflie Zamrod, Mohammed Noor Embi

Research output: Contribution to journalArticle

Abstract

The active part of bacterial toxin enzymes from Burkholderia pseudomallei, Pseudomonas aeruginosa and difteria is ADP-ribosilation domain. This particular domain is conserved among all the three microorganisms. The ADP-ribosilation domain from B. pseudoamllei was PCR amplified using B. pseudomallei virulent genome as a template and primers that were generated based on Pseudomonas aeruginosa ADP-ribosilation sequences. The amplification product was purified and used as a probe (HPCR2) to screen the DNA inserts from B. pseudoamallei that have been cloned into expression vector pSport-I. The objective of this study was to screen eight positive clones obtained ftom early screening using immunoblot and antitoxin generated from rabbit. We also screened three clones that produced negative results during immunoblot screening. The results showed that only one clone (L31) from eight positive clones carried the ADP-ribosilation domain. Manual DNA sequencing of L31 using two primers was able to generate 450bp. Further analysis showed that only one polypeptide from six posibilities was able to be translated without existence of any stop codon.

Original languageUndefined/Unknown
Pages (from-to)101-106
Number of pages6
JournalSains Malaysiana
Volume37
Issue number1
Publication statusPublished - Mar 2008

Fingerprint

Burkholderia pseudomallei
toxins
clones
screening
Pseudomonas aeruginosa
genes
bacterial toxins
antitoxins
stop codon
polypeptides
sequence analysis
rabbits
microorganisms
genome
DNA
enzymes

Keywords

  • ADP-ribosilation domain
  • Burkholderia pseudomallei
  • Manual sequencing

ASJC Scopus subject areas

  • General

Cite this

Penyaringan domain ADP-ribosilasi gen toksin daripada Burkholderia pseudomallei virulen. / Zainal Ariffin, Shahrul Hisham; Mohamed, Rahmah; Zamrod, Zulkeflie; Embi, Mohammed Noor.

In: Sains Malaysiana, Vol. 37, No. 1, 03.2008, p. 101-106.

Research output: Contribution to journalArticle

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N2 - The active part of bacterial toxin enzymes from Burkholderia pseudomallei, Pseudomonas aeruginosa and difteria is ADP-ribosilation domain. This particular domain is conserved among all the three microorganisms. The ADP-ribosilation domain from B. pseudoamllei was PCR amplified using B. pseudomallei virulent genome as a template and primers that were generated based on Pseudomonas aeruginosa ADP-ribosilation sequences. The amplification product was purified and used as a probe (HPCR2) to screen the DNA inserts from B. pseudoamallei that have been cloned into expression vector pSport-I. The objective of this study was to screen eight positive clones obtained ftom early screening using immunoblot and antitoxin generated from rabbit. We also screened three clones that produced negative results during immunoblot screening. The results showed that only one clone (L31) from eight positive clones carried the ADP-ribosilation domain. Manual DNA sequencing of L31 using two primers was able to generate 450bp. Further analysis showed that only one polypeptide from six posibilities was able to be translated without existence of any stop codon.

AB - The active part of bacterial toxin enzymes from Burkholderia pseudomallei, Pseudomonas aeruginosa and difteria is ADP-ribosilation domain. This particular domain is conserved among all the three microorganisms. The ADP-ribosilation domain from B. pseudoamllei was PCR amplified using B. pseudomallei virulent genome as a template and primers that were generated based on Pseudomonas aeruginosa ADP-ribosilation sequences. The amplification product was purified and used as a probe (HPCR2) to screen the DNA inserts from B. pseudoamallei that have been cloned into expression vector pSport-I. The objective of this study was to screen eight positive clones obtained ftom early screening using immunoblot and antitoxin generated from rabbit. We also screened three clones that produced negative results during immunoblot screening. The results showed that only one clone (L31) from eight positive clones carried the ADP-ribosilation domain. Manual DNA sequencing of L31 using two primers was able to generate 450bp. Further analysis showed that only one polypeptide from six posibilities was able to be translated without existence of any stop codon.

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