碩士 / 國立屏東科技大學 / 水產養殖系 / 94 / A coagulation system is essential in both invertebrate and vertebrate animals to prevent excess blood loss from wounds and obstruction of invading microorganisms via the wounds. Transglutaminase (TG) (EC 2.3.2.13) is the only molecule conserved in the coagulation systems of all known invertebrate species. TG cDNA is cloned from the haemocytes of white shrimp (Litopenaeus vannamei) by reverse transcription polymerase chain reaction (RT-PCR) using degenerate primers. The full-length cDNAs of TG consisted of 2624 bp with 2073 bp open reading frame, and encoded 691 amino acids with predicting molecular mass of 77.495 kDa with an estimated pI of 5.73. One putative integrin binding motif (cell adhesion site), RGD (Arg-Gly-Asp) and four calcium binding sites, and one catalytic triad were observed in the TG. The percentage of sequence similarity in L. vannamei TG amino acid sequence identity, other arthropod TG, and human factorXIIIa were about 32.3 to 89.7%. TG activity was detected in all tissues, but no activity was detected in the plasma. This study uses RT-PCR and real-time PCR to analyze the TG mRNA expression in each tissue. The quantitative real-time PCR used to evaluate the effects of the expressions of TG genes in temperature and Vibrio alginolyticus-injections treatments. The results were as follows: TG transcripts in haemocytes were significantly lower at 12 and 24 h in V. alginolyticus-injection treatment, and in hepatopancreas, they increased significantly at 3 h and then recovered to basic value. The TG transcripts expressions in haemocytes at 28 ℃ were higher than 22 and 34 ℃ treatment at 168 h, and the TG transcripts expressions in hepatopancreas at 28 ℃ treatment were significantly higher than 22 and 34 ℃. Rearing at a salinity of 5, 15, 25, 35 and 45 ‰, TG transcripts in haemocytes and hepatopancreas of white shrimp showed no significant difference.
| Identifer | oai:union.ndltd.org:TW/094NPUST086002 |
| Date | January 2006 |
| Creators | Chia-Wen Hung, 洪嘉文 |
| Contributors | Winton Cheng, Mei-Chen Tseng, 鄭文騰, 曾美珍 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 91 |
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