Journal of Animal and Veterinary Advances
ISSN: Online 1993-601XISSN: Print 1680-5593
Abstract
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 ALD gene of Haemonchus contortus (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 Caenorhabditis elegans, Brugia malayi and Onchocerca volvulus. The biochemical assay showed that the recombinant HcALD exhibited enzymatic activity and the optimum temperature and pH for the reaction were 40°C and 7.5, respectively.
How to cite this article:
Ruofeng Yan, Lixin Xu, Jingjing Wang, Xiaokai Song and Xiangrui Li. Cloning and Characterization of Aldolase from Parasitic Nematode Haemonchus contortus.
DOI: https://doi.org/10.36478/javaa.2013.478.486
URL: https://www.makhillpublications.co/view-article/1680-5593/javaa.2013.478.486