TY  - JOUR
T1  - Cloning and Characterization of Aldolase from Parasitic Nematode <I>Haemonchus contortus</I>
AU - Li, Xiangrui AU - Yan, Ruofeng AU - Xu, Lixin AU - Wang, Jingjing AU - Song, Xiaokai 
JO  - Journal of Animal and Veterinary Advances
VL  - 12
IS  - 4
SP  - 478
EP  - 486
PY  - 2013
DA  - 2001/08/19
SN  - 1680-5593
DO  - javaa.2013.478.486
UR  - https://makhillpublications.co/view-article.php?doi=javaa.2013.478.486
KW  - Haemonchus contortus
KW  -aldolase
KW  -RACE
KW  -enzyme activity assay
KW  -region
AB  - Aldolase (ALD) was a glycolytic enzyme which catalyzes the cleavage of fructose 1,6-bisphosphate to glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. In the present research, the <I>ALD</I> gene of <I>Haemonchus contortus</I> (HcALD) was first cloned and characterized. Specific primers for the Rapid Amplification of cDNA Ends (RACE) were designed based on the Expressed Sequence Tag (EST) to amplify the 3' and 5' ends of HcALD. The full length of HcALD cDNA was obtained by overlapping the sequences of 3' and 5' extremities. The Open Reading Frame (ORF) of HcALD was amplification by Reverse Transcription PCR (RT-PCR) and expressed in prokaryotic cell. Then, the biochemical activities of the recombinant HcALD protein were analyzed by assays of enzymatic activity, thermal and pH stabilities. The result showed that the full length cDNA of HcALD was 1235 bp, containing 27 bp of 5' Un-Transcript Region (5' UTR), 1098 bp of ORF and 110 bp of 3' UTR. The deduced amino acid sequence of HcALD was highly similarity to the ALDs from the nematodes <I>Caenorhabditis elegans</I>, <I>Brugia malayi</I> and <I>Onchocerca volvulus</I>. The biochemical assay showed that the recombinant HcALD exhibited enzymatic activity and the optimum temperature and pH for the reaction were 40&deg;C and 7.5, respectively.
ER  -