Abstract
We have investigated the expression of clottable protein (CP) in gonad of tiger shrimp (Penaeus monodon) and extent of its phosphorylation. Polyclonal antibodies against purified CP were prepared from rabbit serum. Using this anti-CP antiserum, the temporal expression of CP in gonads of tiger shrimp was analyzed. It was found that the CP occurs only in mature ovaries but not in immature ovaries and testes. Results of RT-PCR confirmed that these tissues expressed low levels of CP mRNA transcripts. Upon eyestalk-ablation, the ovaries in female shrimps were induced to develop, and the CP expression levels in ovaries were traced chronically by RT-PCR analyses. The expression level peaked on day 3 with an increase of about 40 folds relative to the basal level and returned to normal level (as the control shrimp) at day 12. The shrimp embryos at different intervals from spawning to 16h post-spawning were also collected, and it was found that CP contents were gradually decreased in the embryos until the nauplii were hatched. In addition, purified CP was shown to react with specific anti-phosphoserine, anti-phosphothreonine, and anti-phosphotyrosine antibodies suggesting that CP is a phosphoprotein with all types of phosphorylations. Taken together the results suggest that expression of CP in shrimp ovaries is coupled to ovarian development and CP possibly supply nutrition for shrimp embryo.
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