碩士 / 國立臺灣海洋大學 / 水產養殖學系 / 93 / The outer membrane of gram-negative bacteria secrets endotoxin, which is known to cause sepsis leading to multiple organ failure and even death. The tiger shrimp aquaculture industry has once been developed rapidly in Taiwan, and the success of the industry development has been recognized worldwide. However, the shrimp aquaculture industry has unexpectedly suffered from inability of sustained production, primarily due to serious infection of monodon baculovirus (MBV), Vibrio, and white spot syndrome virus (WSSV). Endotoxin binding proteins reported to date include LALF, LBP, and BPI, which are all high-priced synthetic peptides. It is desirable to search for molecular proteins which are capable to neutralize endotoxin and possess antibiotic functions. For this reason, shrimp anti-lipopolysaccharide factor (SALF) was targeted and cloned in this study. SALF is characterized positive charges at 55-76th amino acids and cysteine domains at both ends. Synthesized csSALF (51-79) is shown effectively to neutralize endotoxin , confirmed by LAL assay. MIC values of SALF against gram-positive bacterias , Micrococcus lutus, Streptococccus bovis, Staphylococcus aureus, and Streptococcus pyognes is 2, 1.25, 1.25, and 1.3 mg/ml, respectively, while those against gram-negative bacterias, Escherichia coli, Klebsiella oxytoca, and Pseudomonas aeruginosa is 1.25, 2.5, and 1.0 mg/ml, respectively. In the MTT assay, both csSALF and lsSALF can suppress hyperplasia of HeLa, MCF-7, and HT-1080 cell lines.SALF expresses lesser in the heart, gill and intestine, but higher in the blood cells, muscle, eyestal, and epithelial tissue, the blood cells in particular. The changes in the gene expression of SALF, peroxinectin, prophenoloxidase and transglutaminase extracted from the tiger shrimp blood cells, were further analyzed by real-time PCR. The results indicated that prophenoloxidase expressed continuously after 12-hr post- stimulation, while other three genes started to express in a large quantity after 72-hr post-stimulation.
| Identifer | oai:union.ndltd.org:TW/093NTOU5086022 |
| Date | January 2005 |
| Creators | Wei-Cheng Liu, 劉韋誠 |
| Contributors | Cheng-Hui Lin, Jyh-Yih Chen, 林正輝, 陳志毅 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 65 |
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