The Tsdaf-21 cDNA was cloned into pET-32a and subsequently expressed in Escherichia coli. The recombinant protein TsDAF-21 was purified and evaluated as an antigen in Western blot. The serum from mouse 14, 21, and 28 days after being infected with Trichinella spiralis recognized rTsDAF-21, but the serum from mouse 7 days post infection of T. spiralis could not react with rTsDAF-21. It implied that the antibody of TsDAF-21 did appear in the host 14 days after infection of T. spiralis. Western blot analysis revealed that the expression of TsDAF-21 in the muscle larvae incubated at room temperature for 8 h and at 37 °C for 4 and 8 h was increased compared to the muscle larvae incubated at 4 °C for 4 and 8 h. The results imply that the expression of TsDAF-21 could be induced at high temperature, not by the cold stress. The expression of TsDAF-21 could be attenuated under the different concentrations of geldanamycin (GA) treatment. Western blot showed that the expression of TsDAF-21 was reduced in the muscle larvae of T. spiralis treated with GA compared to Ampicillin and Gentamycin sulfate treated as control, and this inhibition effect was GA dependent. Other antibiotic treatments did not show any inhibition effects. Immunolocalization showed that TsDAF-21 was expressed in the different stages of T. spiralis including newborn larvae, muscle larvae, and adult worms. TsDAF-21 was mostly localized in nuclei of the cells in the worm. These results suggest that the expression of TsDAF-21 could be induced by the heat stress and attenuated by GA treatment, and TsDAF-21 might be a diagnostic marker for Trichinellosis.
The complete mitochondrial genome sequence of the barred mudskipper Periophthalmus argentilineatus was first determined in this study. The circle genome was 16 509 bp long and consisted of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and 1 control region. The nucleotide composition of the heavy strand of P. argentilineatus is 28.28% for A, 27.83% for C, 16.01% for T, and 27.88% for G, with a slight G + C bias of 55.71%. Only the NADH dehydrogenase subunit 6 (ND6) and eight tRNA genes were encoded on the light strand, other mitochondrial genes were all encoded on the heavy strand. The mitochondrial gene arrangement of P. argentilineatus is similar to those of most other gobies. The phylogenetic analysis using the neighbor-joining method showed that the kinship between Periophthalmus and Boleophthalmus is closer than those between Periophthalmus and other selected genera. Periophthalmus argentilineatus was clustered into one branch with other four species from the same genus Periophthalmus.
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