Gene expression from a cold-treated Swedish isolate of Haemonchus contortus

• Main Author: Martinsson, Daniel
• Format: Others
• Language: Swedish
• Published: Uppsala universitet, Institutionen för medicinsk biokemi och mikrobiologi 2009
• Subjects: Single Strand Hybridization; BigDye Terminator; protein-BLAST; Nematode; Sheep
• Online Access: http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-108579

Totally 84 differentially expressed mRNA clones from infective L3 larvae of the parasite Haemonchus contortus, a blood sucking nematode, were analyzed with single strand hybridization assay (SSH). Altogether 79 clones were sequenced, edited, and compared with proteins found via BLAST in GeneBank. The aim was to investigate gene expression and potential protein expression following storage at 5 °C for 32 weeks. mRNA was extracted from fresh and stored L3. The SSH derived products were cloned into E. coli, purified and sequenced with Big Dye Terminator v3.1, and then compared with uploaded sequences in GeneBank. BLAST showed 59 (70 %) reliable protein results, where 39 (66 %) were of nematode origin.. Three sequences (4 %) were recognized as H. contortus-related metabolic proteins. Further research is necessary to elucidate the role of these proteins in relation to storage. Many of the sequences (36 %) are also present in other nematodes, such as Caenorhabditis briggsae and C. elegans, where whole-genome projects have been conducted. Bigger and more accurate databases need to be developed. Maybe the most significant future project is to sequence the whole genome of H. contortus.