Study of the 49 kDa excretory-secretory protein gene of Trichinella nativa and Trichinella spiralis

Zheng, B.; Xiao, L.; Wang, X.; Li, D.; Lu, Y.; Zhang, Y.; Yan, Q.; Song, M.

doi:10.2478/s11687-007-0018-4

Abstract

To study the function of the 49 kDa excretory-secretory (ES) protein gene (P49) of Trichinella, the genes was amplified by RT-PCR from RNA of Trichinella spiralis and Trichinella nativa and several Chinese Trichinella isolates of domestic animals, and sequenced after being cloned. The amplified products of these parasites produced bands of about 950 bp. The 97.2 % to 100 % nucleotides identity and 94.3 % to 100 % identity of deduced amino acids among P49 gene of these Trichinella strains showed the close relationship of these parasites. The P49 gene of T. nativa was cloned into the BamHI site of the prokaryotic expression vector pET-30a, and the recombinant vector was expressed. The expressed product was 40.8 kDa in size. In Western blot analysis, the expressed product was reactive to sera of mice infected with T. nativa, T. spiralis and their Chinese geographical strains.

References

[1] Bolas-Fernandez, F. (2003): Biological variation in Trichinella species and genotypes. J. Helminthol., 77: 111–118 http://dx.doi.org/10.1079/JOH200317010.1079/JOH2003170
Search in Google Scholar
[2] Dea-Ayuela, M. A., Bolas-Fernandez, F. (1999): Trichinella antigens: a review. Vet Res., 30: 559–571
Search in Google Scholar
[3] Denkers, E. Y., Wassom, D. L., Hayes, C. E. (1990): Characterization of Trichinella spiralis antigens sharing an immunodominant, carbohydrate-associated determinant distinct from phosphorylcholine. Mol. Biochem. Parasitol., 41: 241–250 http://dx.doi.org/10.1016/0166-6851(90)90187-Q10.1016/0166-6851(90)90187-Q
Search in Google Scholar
[4] Despommier, D. D., Muller M. (1976): The stichosome and its secretion granules in the mature muscle larva of Trichinella spiralis. J. Parasitol., 62: 775–785 http://dx.doi.org/10.2307/327896010.2307/3278960
Search in Google Scholar
[5] Gamble, H. R., Anderson, W. R., Graham, C. E., Murrell, K. D. (1983): Diagnosis of swine trichinosis by enzyme-linked immunosorbent assay (ELISA) using an excretory-secretory antigen. Vet. Parasitol., 13: 349–361 http://dx.doi.org/10.1016/0304-4017(83)90051-110.1016/0304-4017(83)90051-1
Search in Google Scholar
[6] Gamble, H. R., Graham, C. E. (1984): Monoclonal antibody-purified antigen for the immunodiagnosis of trichinosis. Am. J. Vet. Res., 45: 67–74
Search in Google Scholar
[7] Gamble, H. R. Murrell, K. D., Marti, H. P. (1986): Inoculation of pigs against Trichinella spiralis using larval excretory-secretory antigens. Am. J. Vet. Res., 47: 2396–2399
Search in Google Scholar
[8] Geerts, S., de Borchgrave, J., Dorny, P., Brandt, J. (2002): Trichinellosis: old facts and new developments. Verh, K. Acad, Geneeskd Belg., 64: 233–248
Search in Google Scholar
[9] Kapel, C. M. (2000): Host diversity and biological characteristics of the Trichinella genotypes and their effect on transmission. Vet. Parasitol., 93: 263–278 http://dx.doi.org/10.1016/S0304-4017(00)00345-910.1016/S0304-4017(00)00345-9
Search in Google Scholar
[10] La Rosa, G., Pozio, E., Rossi, P., Murrell, K. D. (1992): Allozyme analysis of Trichinella isolates from various host species and geographical regions. J. Parasitol., 78: 641–646 http://dx.doi.org/10.2307/328353810.2307/3283538
Search in Google Scholar
[11] La Rosa, G., Marucci, G., Pozio, E. (2003): Biochemical analysis of encapsulated and non-encapsulated species of Trichinella (Nematoda, Trichinellidae) from cold- and warm-blooded animals reveals a high genetic divergence in the genus. Parasitol. Res., 91: 462–466 http://dx.doi.org/10.1007/s00436-003-0981-810.1007/s00436-003-0981-814557876
Search in Google Scholar
[12] Li, A., Yuan, H. (1989): Experimental Studies on Functional Antigens of Larvae of Trichinella Spiralis in Muscle (in Chinese, with English abstract). Bull. Vet. Coll. Peoples Liberation Army, 9: 270–273
Search in Google Scholar
[13] Lichtenfels, J. R., Murrell, K. D., Pilitt, P. A. (1983): Comparison of Three Subspecies of Trichinella spiralis by Scanning Electron Microscopy. J. Parasitol., 69: 1131–1140 http://dx.doi.org/10.2307/328087810.2307/3280878
Search in Google Scholar
[14] Liu, M. Y., Xu, K. C., Song, M.X., Zhu, X.P., An, C. L., Hou, S. L., Zhang, R. T., Boireau, P., Fabien, J. F., Trap, C., Vayssire, M., Vallet, C., Perret, C., Soule, C. (1998): Further species identification of Trichinella isolate SW of Chinese Cat (in Chinese, with English abstract). Chin. J. Vet. Sci. Technol., 28: 8–10
Search in Google Scholar
[15] Nagano, I., Wu, Z., Nakada, T., Matsuo, A., Takahashi, Y. (2001): Molecular cloning and characteri-zation of a 21 kDa protein secreted from Trichinella pseudospiralis. J Helminthol., 75: 273–278
Search in Google Scholar
[16] Oivanen, L., Kapel, C. M., Pozio, E., La Rosa, G., Mikkonen, T., Sukura, A. (2002): Associations between Trichinella species and host species in Finland. J. Parasitol., 88: 84–88
Search in Google Scholar
[17] Pozio, E., La Rosa, G., Rossi, P., Murrell, K. D. (1992a): Biological characterization of Trichinella from various host species and geographical regions. J. Parasitol., 78: 647–653 http://dx.doi.org/10.2307/328353910.2307/3283539
Search in Google Scholar
[18] Pozio, E., La Rosa, G., Murrell, K.D., Lichtenfels, J. R. (1992b): Taxonomic revision of the genus Trichinella. J. Parasitol., 78: 654–659 http://dx.doi.org/10.2307/328354010.2307/3283540
Search in Google Scholar
[19] Pozio, E. Foggin, C. M., Marucci, G., La Rosa, G., Sacchi, L., Corona, S., Rossi, P., Mukaratirwa S. (2002): Trichinella zimbabwensis n. sp. (Nematoda), A new nonencapsulated species from crocodiles (Crocodylus niloticus) in Zimbabwe also infecting mammals. Int. J. Parasitol., 32: 1787–1799 http://dx.doi.org/10.1016/S0020-7519(02)00139-X10.1016/S0020-7519(02)00139-X
Search in Google Scholar
[20] Sambrook, J., Fritsch, E. F., Maniatis, T. (1999): Translated by Jin D.Y., Li, M.F. et al.. 2nd (Ed.): Molecular Cloning: A Laboratory Manual (in Chinese). Science Press, Beijing, China: 16–898
Search in Google Scholar
[21] Silberstein, D. S., Despommier, D. D. (1984): Antigens from Trichinella spiralis that induce a protective response in the mouse. J. Immunol., 132: 898–904
Search in Google Scholar
[22] Smith, H. J. (1985): Infectivity of Canadian isolates of Trichinella spiralis nativa for swine, rats and carnivores. Can. J. Comp. Med., 49: 88–90
Search in Google Scholar
[23] Sofronic-Milosavljevic, L., Pozio, E., Patrascu, I.V., Skerovic, N., Gomez Morales, M. A., Gamble, H. R. (2001): Immunodiagnosis of Trichinella infection in the horse. Parasite, 8: 260–262 10.1051/parasite/200108s2260
Search in Google Scholar
[24] Song, M. X., Zhou, Y. C., Li, S. S. (1994): Comparative study on isozyme patterns of Trichinella SP. From swine and dogs in harbin region, China (in Chinese, with English abstract). J. Northeast Agr. Univ., 25: 260–265
Search in Google Scholar
[25] Song, M. X., Zhou, Y. C., Wang, H. F. (1998): Infectivity of Trichinella SP. from swine for swine and dogs in Harbin region, China (in Chinese, with English abstract). A. Vet. Zootech. Sinica, 29: 267–270
Search in Google Scholar
[26] Su, X. Z., Prestwood, A. K. (1991a): A dot-ELISA mimicry western blot test for the detection of swine trichinellosis. J. Parasitol., 77: 76–82 http://dx.doi.org/10.2307/328256110.2307/3282561
Search in Google Scholar
[27] Su, X. Z., Prestwood, A. K., McGraw, R. A. (1991b): Cloning and expression of complementary DNA encoding an antigen of Trichinella spiralis. Mol. Biochem. Parasitol., 45: 331–336 http://dx.doi.org/10.1016/0166-6851(91)90101-B10.1016/0166-6851(91)90101-B
Search in Google Scholar
[28] Sukura, A, Nareaho, A, Mikkonen, T, Niemi, M, Oivanen, L. (2002): Trichinella nativa and T. spiralis induce distinguishable histopathologic and humoral responses in the raccoon dog (Nyctereutes procyonoides). Vet. Pathol., 39: 257–265 http://dx.doi.org/10.1354/vp.39-2-25710.1354/vp.39-2-257
Search in Google Scholar
[29] Turčeková, L.’, Borošková, Z., Tomašovičová, O., Reiterová, K., Kinčeková, J. (1997): Immunochemical analysis of larval antigens of Trichinella spiralis and T. pseudospiralis. Helminthologia, 34: 241–243
Search in Google Scholar
[30] Vassilatis, D. K., Despommier, D., Misek, D. E., Polvere, R. I., Gold, A. M., Lex, H. T. Van der Ploeg (1992): Analysis of a 43-kDa glycoprotein from the intracellular parasitic nematode Trichinella spiralis. J. Biol. Chem., 267: 18459–18465
Search in Google Scholar
[31] Wisnewski, N., McNeil, M., Grieve, R. B., Wassom, D. L. (1993): Characterization of novel fucosyl- and tyvelosyl-containing glycoconjugates from Trichinella spiralis muscle stage larvae. Mol. Biochem. Parasitol., 61: 25–36 http://dx.doi.org/10.1016/0166-6851(93)90155-Q10.1016/0166-6851(93)90155-Q
Search in Google Scholar
[32] Wu, Z., Nagano, I., Takahashy, Y. (1998): The detection of Trichinella with polymerase chain reaction (PCR) primers constructed using sequences of random amplified polymorphic DNA (RAPD) or sequences of complementary DNA encoding excretory-secretory (E-S) glycoproteins. Parasitology, 117: 173–183 http://dx.doi.org/10.1017/S003118209800288110.1017/S0031182098002881
Search in Google Scholar
[33] Wu, Z., Nagano, I., Pozio, E., Takahashy, Y. (1999): Polymerase chain reaction-restriction fragment length polymerphism (PCR-RAPD) for the identification of Trichinella isolates. Parasitology, 118: 211–218 http://dx.doi.org/10.1017/S003118209800367910.1017/S0031182098003679
Search in Google Scholar
[34] Yan, Y. H., Tong, G. Z., Lu, J. L. (1997): Cloning of two structural genes encoding specific proteinsin ES antigen from Trichinella spiralis muscle larvae and their high level expression in E. coli and yeast (in Chinese, with English abstract). Chin. J. Vet. Sci., 17: 581–588
Search in Google Scholar
[35] Yao, C., McGraw, R. A., Prestwood, A. K. (1997): A complementary DNA encoding an antigen from Trichinella spiralis muscle larvae and its analog from Trichinella T5 of bobcat origin: sequence, cloning and expressions. J. Parasitol., 27: 425–430 http://dx.doi.org/10.1016/S0020-7519(97)00001-510.1016/S0020-7519(97)00001-5
Search in Google Scholar
[36] Zarlenga, D. S., Gamble, H. R. (1990): Molecular cloning and expression of an immunodominant 53kDa Excretory-Secretory antigen from Trichinella spiralis muscle larvae. Mol. Biochem Parasitol., 42: 165–174 http://dx.doi.org/10.1016/0166-6851(90)90159-J10.1016/0166-6851(90)90159-J
Search in Google Scholar
[37] Zarlenga, D.S., La Rosa, G. (2000): Molecular and biological methods for parasite differentiation within the genus Trichinella. Vet. Parasitol., 93: 279–292 http://dx.doi.org/10.1016/S0304-4017(00)00346-010.1016/S0304-4017(00)00346-0
Search in Google Scholar
[38] Zhu, X. P., Liu, M. Y., Wang, F.Y., Zhou, L., Song, M. X. (1998): Identification of three isolates of Trichinella in China on the basis of restriction fragment length polymerphisms (RFLP) analysis (in Chinese, with English abstract). Acta. Parasitol. Med. Entomol. Sin, 5: 101–105
Search in Google Scholar

Study of the 49 kDa excretory-secretory protein gene of Trichinella nativa and Trichinella spiralis

Abstract

Paradigm

My account