碩士 / 國立屏東科技大學 / 水產養殖系 / 93 / This research is aimed to comprehend the molecular characteristic of the cell adhesion protein, peroxinectin in freshwater giant prawn, Macrobrachium rosenbergii, through molecular cloning of peroxinectin cDNA and its expression. The profile of peroxinection expression under different ontogenetic development and molt stages, sodium alginate-fed, and treatments of various bactericides, such as sulfate, B.K.C., and trichlorfon.
Expression of peroxinectin cDNA was determined from haemocytes of the giant freshwater prawn, Macrobrachium rosenbergii, by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA using oligonucleotide primers based on the peroxinectin sequence of white shrimp, Litopenaeus vannamei; tiger shrimp, Penaeus monodon; and freshwater crayfish, Pacifastacus leniusculus. The full-length cDNAs of peroxinectin consisted of 2403 bp open reading frame encode 801 amino acids (aa) with predicted include a 20-aa signal peptide, molecular mass of the mature protein (781 aa) was 88.7 kDa with an
estimated pI of 6.8. A putative peroxidase domain (~500 amion acid) and a putative integrin-binding motif, KGD (Lys-Gly-Asp), was observed in prawn peroxinectin at the C-terminal. Sequence comparison showed that peroxinectin-deduced amino acids of M. rosenbergii had overall similarities of 64%, 62%, and 66% to those of P. monodon, P. leniusculus, and L. vannamei, respectively. To evaluate exogenous and endogenous factors that affect expressions of peroxinectin genes by quantitative real-time PCR, the results were showed: Ontogenic expression of peroxinectin was studied by RT-PCR that peroxinectin mRNA expressed endogenously by the embryo from the sixth days after fertilization to hatching out. In different moult stages, peroxinectin gene expression at A stage were significantly higher than the other stage (B, C, D0/D1, D2/D3). The expressions of peroxinectin were more down-regulated after one-week treatment of containing foods 2000 mg/kg diet sodium alginate than that of 1000 mg/kg diet ones. Peroxinectin genes expression was significantly decreased after carbon powder administration, dead Lactococcus garvieae injection, or live L. garvieae injection at 3 h and recovery was delayed in prawns injected with live L. garvieae. Peroxinectin genes expression was significantly decreased for the prawn exposed to 0.1, 0.2, 0.3 and 0.4 mg/l cupper sulfate, 0.6 and 1 mg/l B.K.C., 0.3, 0.6 and 1 mg/l trichlorfon for 96h. The expression of peroxinectin were obviously influenced by the change of internal and external environmental factors, probably through the proPO system that resulted in the change in immune function and disease tolerance.
| Identifer | oai:union.ndltd.org:TW/093NPUST086005 |
| Date | January 2005 |
| Creators | Pei-Yi Hsu, 許蓓怡 |
| Contributors | Winton Cheng, 鄭文騰 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 89 |
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