Cloning and expression of a TCTP homolog from the ovaries of banana prawn
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Ovarian development in prawns is not well understood. The up-regulated prawn ovarian translationally controlled tumor protein (Pmer-TCTP) of the banana prawn (Penaeus merguiensis) was quantitated by real-time PCR. The results showed that the gene has a significantly increased expression (P < 0.05) in the early stage of matured ovaries (stage 1) whereas Pmer-TCTP transcripts were not significantly increased at the later stage (stages 2 and 3) when compared to the non-matured ovaries. This result corresponds to the result of the western blot analysis when compared with the amounts of native Pmer-TCTP protein from different vitellogenic stages. The result of the western blot analysis indicated that the native Pmer-TCTP is present in higher concentration at the early stage and the molecular mass of the native Pmer-TCTP was about 26 kDa. The deduced amino acid sequence of Pmer-recombinant TCTP was found to be 98% (4e-91) similar to Penaeus monodon TCTP (Pm-TCTP), 44% (3e−27) to human TCTP and 42% (2e−25) to mouse TCTP. The expressed Pmer-recombinant TCTP in Escherichia coli produced a protein of about 19.2 kDa on SDS-PAGE and both the native Pmer-TCTP and Pmer-recombinant TCTP interacted with elongation factor 1 alpha.
KeywordsTranslationally Control Tumor Protein Suppressive Subtractive Hybridization Control Tumor Protein Vitellogenic Stage Myosin Light Chain Kinase Activity
We are most grateful to the Trang Coastal Aquaculture Station, Trang province, Thailand, for the prawn samples, Penaeus merguiensis. This work was supported by the Royal Golden Jubilee Graduate Program from the Thailand Research Fund (TRF) for Wiriya Loongyai (4.C.PS/45/G.1) as well as the Excellent Scholarship in Biochemistry of Prince of Songkla University. We thank Dr. Brian Hodgson, Faculty of Science, Prince of Songkla University, for reading the manuscript and for valuable comments.
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