Hostname: page-component-78c5997874-g7gxr Total loading time: 0 Render date: 2024-11-16T03:25:11.315Z Has data issue: false hasContentIssue false
  • English
  • Français

An evaluation of serial analysis of gene expression (SAGE) in the parasitic nematode, Haemonchus contortus

Published online by Cambridge University Press:  06 January 2005

P. J. SKUCE ,
R. YAGA ,
F. A. LAINSON  and
D. P. KNOX
P. J. SKUCE
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, Scotland, UK
R. YAGA
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, Scotland, UK
F. A. LAINSON
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, Scotland, UK
D. P. KNOX
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, Scotland, UK
Get access Rights & Permissions [Opens in a new window]

Abstract

This study evaluated a relatively new molecular technique, serial analysis of gene expression (SAGE), as a tool for quantifying gene expression in the ovine abomasal nematode Haemonchus contortus for which there is relatively limited (~20% gene coverage) sequence information. SAGE technology generates data that are both qualitative and quantitative and, as such, compliments other functional genomics approaches such as EST analysis and micro-array. Prior to embarking on large-scale comparisons, the present study was initiated to establish (i) how well SAGE and EST data taken from the same life-cycle stage would compare, (ii) how easily SAGE tags could be assigned to genes given that the genome sequence is not available and (iii) whether it would be possible to extend the sequences of the SAGE tags to facilitate their identification. Of 2825 tag sequences analysed from adults harvested 28 days post-infection, the identity of the encoding gene could be ascribed to 63% of the tags. The relative abundance of these genes, arbitrarily categorized on the basis of function, was comparable with that of an EST dataset also from adults (n=2317). In addition, tag sequences could be readily extended and thereby identified using a tag-based primer and Reverse Transcription-PCR.

Keywords

Haemonchus contortusserial analysis of gene expressionSAGE
Type
Research Article
Information
Parasitology , Volume 130 , Issue 5 , May 2005 , pp. 553 - 559
Copyright
© 2005 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

THE C. ELEGANS SEQUENCING CONSORTIUM. ( 1998). Genome sequence of the nematode C. elegans: a platform for investigating biology. Science 282, 2012–2018.
HOEKSTRA, R., VISSER, A., OTSEN, M., TIBBEN, J., LENSTRA, J. A. & ROOS, M. H. ( 2000). EST sequencing of the parasitic nematode Haemonchus contortus suggests a shift in gene expression during transition to the parasitic stages. Molecular and Biochemical Parasitology 110, 53–68.CrossRefGoogle Scholar
HARTMAN, D., DONALD, D. R., NIKOLAOU, S., SAVIN, K. W., HASSE, D., PRESIDENTE, P. J. & NEWTON, S. E. ( 2001). Analysis of developmentally regulated genes of the parasite Haemonchus contortus. International Journal for Parasitology 31, 1236–1245.CrossRefGoogle Scholar
LONGBOTTOM, D., REDMOND, D. L., RUSSELL, M., LIDDELL, S., SMITH, W. D. & KNOX, D. P. ( 1997). Molecular cloning and characterisation of a putative aspartate proteinase associated with a gut membrane protein complex from adult Haemonchus contortus. Molecular and Biochemical Parasitology 88, 63–72.CrossRefGoogle Scholar
JONES, S. J., RIDDLE, D. L., POUZYREV, A. T., VELCULESCU, V. E., HILLIER, L., EDDY, S. R., STRICKLIN, S. L., BAILLIE, D. L., WATERSTON, R. & MARRA, M. A. ( 2001). Changes in gene expression associated with developmental arrest and longevity in Caenorhabditis elegans. Genome Research 11, 1323–1324.CrossRefGoogle Scholar
KNOX, D. P. ( 2004). Technological advances and genomics in metazoan parasites. International Journal for Parasitology 34, 139–152.CrossRefGoogle Scholar
KNOX, D. P., REDMOND, D. L., NEWLANDS, G. F., SKUCE, P. J., PETTIT, D. & SMITH, W. D. ( 2003). The nature and prospects for gut membrane proteins as vaccine candidates for Haemonchus contortus and other ruminant trichostrongyloids. International Journal for Parasitology 33, 1129–1137.CrossRefGoogle Scholar
MUNASINGHE, A., PATANKAR, S., COOK, B. P., MADDEN, S. L., MARTIN, R. K., KYLE, D. E., SHOAIBI, A., CUMMINGS, L. M. & WIRTH, D. F. ( 2001). Serial analysis of gene expression (SAGE) in Plasmodium falciparum: application of the technique to A-T rich genomes. Molecular and Biochemical Parasitology 113, 23–34.CrossRefGoogle Scholar
NOWAK, R. ( 1995). Entering the post genome era. Science 270, 368–371.CrossRefGoogle Scholar
PARKINSON, J., MITREVA, M., HALL, N., BLAXTER, M. & McCARTER, J. P. ( 2003). 400000 nematode ESTs on the Net. Trends in Parasitology 19, 283–286.CrossRefGoogle Scholar
SKUCE, P. J., REDMOND, D. L., LIDDELL, S., STEWART, E. M., SMITH, W. D. & KNOX, D. P. ( 1999). Molecular cloning and characterisation of gut-derived cysteine proteinases associated with a host protective extract from Haemonchus contortus. Parasitology 119, 396–405.Google Scholar
SKUCE, P. J., NEWLANDS, G. F. J., STEWART, E. M., PETTIT, D., SMITH, S. K., SMITH, W. D. & KNOX, D. P. ( 2001). Cloning and characterisation of thrombospondin, a novel multidomain glycoprotein found in association with a host protective gut extract from Haemonchus contortus. Molecular and Biochemical Parasitology 117, 241–244.CrossRefGoogle Scholar
SMITH, W. D., SMITH, S. K., PETTIT, D., NEWLANDS, G. F. & SKUCE, P. J. ( 2000). Relative protective properties of three membrane glycoprotein fractions from Haemonchus contortus. Parasite Immunology 22, 63–71.CrossRefGoogle Scholar
TUTEJA, R. & TUTEJA, N. ( 2004). Serial analysis of gene expression: applications in human studies. Journal of Biomedicine and Biotechnology 2004, 113–120.CrossRefGoogle Scholar
VAN DEN BERG, A., VAN DER LEIJ, J. & POPPEMA, S. ( 1999). Serial analysis of gene expression: rapid RT-PCR analysis of unknown SAGE tags. Nucleic Acids Research 27, 17.CrossRefGoogle Scholar
VELCULESCU, V. E., ZHANG, L., VOGELSTEIN, B. & KINZLER, K. W. ( 1995). Serial analysis of gene expression. Science 270, 484–487.CrossRefGoogle Scholar
YAMAMOTO, M., WAKATSUKI, T., HADA, A. & RYO, A. ( 2001). Use of serial analysis of gene expression (SAGE) technology. Journal of Immunological Methods 250, 45–66.CrossRefGoogle Scholar
YE, S. Q., ZHANG, L. Q., ZHENG, F., VIRGIL, D. & KWITEROVICH, P. O. ( 2000). MiniSAGE: Gene expression profiling using serial analysis of gene expression from 1 μg total RNA. Analytical Biochemistry 287, 144–152.CrossRefGoogle Scholar