碩士 / 國立屏東科技大學 / 水產養殖系所 / 96 / A coagulation system is essential in both invertebrate and vertebrate animals to prevent excess blood loss from wounds and also obstruct invasion of 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 giant freshwater prawn (Macrobrachium rosenbergii) by reverse transcription polymerase chain reaction (RT-PCR) using degenerate primers. The full-length cDNA of TG consisted of 2751 bp with 2334 bp open reading frame, and encoded 778 amino acids with predicting molecular mass of 87.67 kDa with an estimated pI of 5.40. Two putative integrin binding motif (cell adhesion site) RGD (Arg-Gly-Asp) were observed in the TG. Neither a typical hydrophobic leader sequence nor a transmembrane domain could be identified from the deduced sequence, Thus, the TG may be a typical cytoplasmic protein. The TG sequence was similar to crayfish and other invertebrates TG sequences (56%~69%). TG activity was detected in all tissues, except plasma. Transcription of TG mRNA by RT-PCR were mainly detected in hemocytes, testis and ovary. Real-time RCR was used to analyze the effects of TG mRNA expression in different moult stage and Lactococcus garvieae injection treatments. The TG transcript and activity decreased with the increased of coagulation time significantly after 3 h challenged by L. garvieae. Later on, the mRNA and TG activity increased simultaneously with the decreased of coagulation time after 12 h challenged by L. garvieae. TG transcripts of moult stage A showed the lowest level as well as TG activity.
Keywords: Macrobrachium rosenbergii, Transglutaminase, typical cytoplasmic protein, Integrin binding site, Real-time RCR
| Identifer | oai:union.ndltd.org:TW/096NPUS5086005 |
| Date | January 2008 |
| Creators | Yun Chih Chiu, 邱允志 |
| Contributors | Winton Cheng, Chun Hung Liu, 鄭文騰, 劉俊宏 |
| 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 |
Page generated in 0.0012 seconds