Molecular cloning and characterisation of a venom allergen AG5-like cDNA from Meloidogyne incognita.
RNA fingerprinting was used to identify RNAs that were expressed in parasitic second-stage juveniles of Meloidogyne incognita, but absent from or reduced in preparasitic second-stage juveniles. A cDNA encoding a putative secretory protein was cloned from a M. incognita second-stage juvenile cDNA library by probing with a 0.5kb fragment derived from fingerprinting that was more strongly expressed in parasitic second-stage juveniles. The cDNA, named Mi-msp-1, contained an open reading frame encoding 231 amino acids, with the first 21 amino acids being a putative secretory signal. In Southern blot analysis the Mi-msp-1 hybridised with genomic DNA from M. incognita, Meloidogyne arenaria, Meloidogyne javanica, but not Meloidogyne hapla, Heterodera glycines or Caenorhabditis elegans. In Northern blot analysis a 1kb transcript was detected in both preparasitic and parasitic second-stage juveniles, but not in adult females of M. incognita. Comparing the predicted amino acid sequence with protein databases revealed significant similarity to the venom allergen antigen 5 family of proteins in hymenoptera insects and homologues found in several other nematode species.
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Department of Plant Pathology, University of Georgia, Athens, GA 30603-7274, USA.
Shields
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J
Allen
R
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Hussey
R S
RS
D000485:Allergens / Q000235:genetics* / Q000276:immunology; D000818:Animals; D000947:Antigens, Helminth; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018509:DNA, Helminth; D015801:Helminth Proteins; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000469:parasitology*; D014431:Tylenchoidea / Q000235:genetics* / Q000276:immunology; D014688:Venoms / Q000235:genetics* / Q000276:immunology
D000485:Allergens; D000947:Antigens, Helminth; D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D014688:Venoms; C407782:msp-1 allergen, Meloidogyne