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Detection Of Free Living Amoebae, Acanthamoeba And Naegleria, In Swimming Pools, Malaysia, Init Ithoi Dec 2010

Detection Of Free Living Amoebae, Acanthamoeba And Naegleria, In Swimming Pools, Malaysia, Init Ithoi

Init Ithoi

This study reports the detection of Acanthamoeba and Naegleria species in 14 swimming pools around Petaling Jaya and Kuala Lumpur, Malaysia. Sampling was carried out at 4 sites (the platforms (P), wall (W), 1 meter from the wall (1) and middle (2)) of each swimming pool. These free living amoebae (FLA) were detected under light and inverted microscopes after being cultured on the surface of non-nutrient agar lawned with Escherichia coli. Acanthamoeba species were detected in higher number of culture plates from all sampling sites of all the swimming pools. While Naegleria, were detected in fewer culture plates at 3 …


Cloning And Expression Of Toxoplasma Gondii Dense Granular Protein 4 (Gra4) In Pichia Pastoris, Init Ithoi Dec 2010

Cloning And Expression Of Toxoplasma Gondii Dense Granular Protein 4 (Gra4) In Pichia Pastoris, Init Ithoi

Init Ithoi

GRA4 of Toxoplasma gondii has been shown to prompt IgG, IgM and IgA responses in previous studies and is thus considered one of the major immunogenic proteins from T. gondii that can be used for both diagnostics purposes and vaccine development. This study seeks to clone and express the GRA4 in Pichia pastoris, which has numerous advantages over other systems for expression of eukaryotic proteins. In order to achieve this, the gene was cloned into the pPICZ alpha A expression vector, which was then incorporated into the P. pastoris genome via insertional integration for expression of the recombinant protein, under …


Optimization For High-Level Expression In Pichia Pastoris And Purification Of Truncated And Full Length Recombinant Sag2 Of Toxoplasma Gondii For Diagnostic Use, Init Ithoi May 2010

Optimization For High-Level Expression In Pichia Pastoris And Purification Of Truncated And Full Length Recombinant Sag2 Of Toxoplasma Gondii For Diagnostic Use, Init Ithoi

Init Ithoi

SAG2 is one of the major surface antigens of the intracellular protozoan parasite Toxoplasma gondii. In the present study, truncated recombinant SAG2(S) and full length recombinant SAG2(T) of T. gondii were optimally produced (similar to 15 mg/liter) in Pichia pus tons expression system using BMMY medium at pH 3, 25 degrees C in 0.5-1% methanol and a time-course of 1-2 days. The recombinant proteins were purified using a commercial gel filtration purification system obtaining similar to 33% recovery. The purified SAG2(S) and SAG2(T) showed molecular masses of 45 and 36 kDa by SDS-PAGE, respectively. The recombinant proteins were evaluated by …