摘要
Trichinella spiralis, an intracellular parasitic nematode, can cause severe foodborne zoonosis, trichinellosis. The life cycle of T. spiralis consists of adult(Ad), muscle larvae(ML) and newborn larvae(NBL). The protein profiles in different developmental stages of the parasite remain unknown. In the present study, proteins from lysates of Ad, ML and NBL were identified by isobaric tags for relative and absolute quantitation(iTRAQ). A total of 4,691 proteins were identified in all the developmental stages, of which 1,067 proteins were differentially expressed(DEPs). DEPs in the three developmental stages of T. spiralis were involved in diverse functions in the life cycle, including regulation of worm growth and sexual maturity(e.g. major sperm protein), metabolism and utilization of carbohydrates, lipids and nucleotides(e.g. glucose-6-phosphate isomerase), modulation of host cell functions for the parasite benefit via excretory-secretory(E-S) antigens(e.g. serine protease), and other crucial developmental processes that occur at distinct stages. The number of up-regulated proteins in NBL was higher than that of the other two groups. The protein profiles from Ad, ML and NBL were compared in pairs. The identified proteins were involved in various functions of T. spiralis life cycle, including sexual maturity, metabolism, utilization of carbohydrates, lipids and nucleotides, and other crucial developmental processes that occur at distinct stages. The possible functions of the identified proteins were predicted by the gene function classification system GO, which has three ontologies including molecular function(16,015 proteins), cellular component(8,740 proteins) and biological process(4,676 proteins). Eight genes from DEPs designated E5S8I3 and E5SED3(major sperm protein, gene locus tag, Tsp_00052 and Tsp_02102), B7SIW4(serine protease gene locus tag, Tsp_SP-1.2), Q27071(53kD a E-S antigen, gene locus tag, Tsp_gp53), E5SF13(zinc finger protein, gene locus tag Tsp_02330), E5RY73(histone H2 B subunit, gene locus tag, Tsp_07127), E5S882(DNA polymerase ε subunit, gene locus tag, Tsp_07920), and E5S6R9(neural proliferation differentiation and control protein, gene locus tag, Tsp_07015) were detected using quantitative RT-PCR, which were showed identical results to the iTRAQ analysis. The identification of these proteins in NBL, ML and Ad3 has facilitated a better understanding of the developmental mechanism of T. spiralis and the host-parasite interactions.
Trichinella spiralis, an intracellular parasitic nematode, can cause severe foodborne zoonosis, trichinellosis. The life cycle of T. spiralis consists of adult(Ad), muscle larvae(ML) and newborn larvae(NBL). The protein profiles in different developmental stages of the parasite remain unknown. In the present study, proteins from lysates of Ad, ML and NBL were identified by isobaric tags for relative and absolute quantitation(iTRAQ). A total of 4,691 proteins were identified in all the developmental stages, of which 1,067 proteins were differentially expressed(DEPs). DEPs in the three developmental stages of T. spiralis were involved in diverse functions in the life cycle, including regulation of worm growth and sexual maturity(e.g. major sperm protein), metabolism and utilization of carbohydrates, lipids and nucleotides(e.g. glucose-6-phosphate isomerase), modulation of host cell functions for the parasite benefit via excretory-secretory(E-S) antigens(e.g. serine protease), and other crucial developmental processes that occur at distinct stages. The number of up-regulated proteins in NBL was higher than that of the other two groups. The protein profiles from Ad, ML and NBL were compared in pairs. The identified proteins were involved in various functions of T. spiralis life cycle, including sexual maturity, metabolism, utilization of carbohydrates, lipids and nucleotides, and other crucial developmental processes that occur at distinct stages. The possible functions of the identified proteins were predicted by the gene function classification system GO, which has three ontologies including molecular function(16,015 proteins), cellular component(8,740 proteins) and biological process(4,676 proteins). Eight genes from DEPs designated E5S8I3 and E5SED3(major sperm protein, gene locus tag, Tsp_00052 and Tsp_02102), B7SIW4(serine protease gene locus tag, Tsp_SP-1.2), Q27071(53kD a E-S antigen, gene locus tag, Tsp_gp53), E5SF13(zinc finger protein, gene locus tag Tsp_02330), E5RY73(histone H2 B subunit, gene locus tag, Tsp_07127), E5S882(DNA polymerase ε subunit, gene locus tag, Tsp_07920), and E5S6R9(neural proliferation differentiation and control protein, gene locus tag, Tsp_07015) were detected using quantitative RT-PCR, which were showed identical results to the iTRAQ analysis. The identification of these proteins in NBL, ML and Ad3 has facilitated a better understanding of the developmental mechanism of T. spiralis and the host-parasite interactions.
引文
