PMID- 23959483
OWN - NLM
STAT- MEDLINE
DCOM- 20140310
LR  - 20130820
IS  - 2521-9855 (Electronic)
IS  - 0127-5720 (Linking)
VI  - 30
IP  - 2
DP  - 2013 Jun
TI  - Heterologous expression of uncharacterized parasitic nematode-specific growth 
      factor-like protein of Setaria digitata in Pichia pastoris expression systems.
PG  - 181-92
AB  - Setaria digitata is an animal filarial parasite infecting cattle and other 
      ungulates causing mild to severe diseases, and infection of human by this 
      nematode has also been reported. Therefore, this study was undertaken as an 
      initial step towards the characterization of an uncharacterized gene of S. 
      digitata, that encode for parasitic nematode-specific growth factor-like protein 
      to which Wucheraria bancrofti and other filarial and parasitic nematodes have 
      homologous counterparts (revealed by bioinformatic analyses), by means of protein 
      expression and purification in Pichia pastoris. In Pichia expression system, S. 
      digitata uncharacterized protein (SDUP) was expressed both as secretory and 
      intracellular protein. SDS-PAGE and western blot analyses revealed expression of 
      recombinant SDUP in P. pastoris, showing a band of 23.8 kDa, which is similar to 
      theoretical molecular weight and also this protein was shown to be more 
      concentrated into insoluble fraction than soluble fraction of cell lysates 
      suggesting that the protein to be more hydrophobic than hydrophilic. Recombinant 
      SDUP was successfully purified using Ni-NTA affinity chromatography purification 
      technique and three dimensional structural studies of the purified protein are 
      currently being progressed. This is the first report of the expression of S. 
      digitata protein in P. pastoris.
FAU - Rodrigo, W W
AU  - Rodrigo WW
AD  - Department of Chemistry, Faculty of Science, University of Colombo, 90, 
      Cumaratunga Munidasa Mawatha, Colombo 03, Sri Lanka.
FAU - Dassanayake, R S
AU  - Dassanayake RS
FAU - Weerasena, S J
AU  - Weerasena SJ
FAU - Karunanayake, E H
AU  - Karunanayake EH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Malaysia
TA  - Trop Biomed
JT  - Tropical biomedicine
JID - 8507086
RN  - 0 (Helminth Proteins)
RN  - 0 (Intercellular Signaling Peptides and Proteins)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Animals
MH  - Cattle
MH  - Chromatography, Affinity
MH  - Cloning, Molecular
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Filarioidea/*genetics
MH  - *Gene Expression
MH  - Helminth Proteins/*biosynthesis/chemistry/genetics/isolation & purification
MH  - Hydrophobic and Hydrophilic Interactions
MH  - Intercellular Signaling Peptides and 
      Proteins/*biosynthesis/chemistry/genetics/isolation & purification
MH  - Molecular Weight
MH  - Pichia/genetics
MH  - Recombinant Proteins/biosynthesis/chemistry/genetics/isolation & purification
EDAT- 2013/08/21 06:00
MHDA- 2014/03/13 06:00
CRDT- 2013/08/21 06:00
PHST- 2013/08/21 06:00 [entrez]
PHST- 2013/08/21 06:00 [pubmed]
PHST- 2014/03/13 06:00 [medline]
PST - ppublish
SO  - Trop Biomed. 2013 Jun;30(2):181-92.