碩士 / 國立臺灣海洋大學 / 水產養殖學系 / 94 / Innate immune system of fish includes antimicrobial and antivirus genes. MX gene is interferon inducible gene. And it’s functions mainly through preventing dsRNA virus from replicating. Antimicrobial peptide (AMP) that can be found among different kinds of organism, plays important roles on first line of defense of pathogen infection. AMP is a physiologic barriers of innate immune. recently, grouper larva was seriously infected by virus and bacteria, hence manipulate of MX and AMP might be a key in solving these problems. Therefore, studying of innate immune system is very important for grouper. In the present study, degenerate primers are designed base on known MX, AMP amino acids and nucleic acids sequences. Both MX and AMP genes cDNA are isolated from giant grouper (Epinephelus lanceolatus) gills for full length cDNA were cloned by using RT-PCR and 5’,3’RACE technique. MX gene cDNA consists of 1881 base pairs and can be deduced into 626 amino acids. Meanwhile, AMP gene cDNA consists of 494 base pairs and can be deduced into 67 amino acids. Both genes express in multiple tissues especially in gills. Real-time quantitative PCR results reveal that changes in salinity have significantly increased MX gene expression. Furthermore, poly I:C injections also increase MX gene expression, regardless of concentration or duration of times. However, no significant changes in the expression of AMP after poly I:C (polyinosinic-polycytidylic acid) injection. Expression of MX genes arise remarkable by immersion of Chinese herbs i.e.Isatis indigotica, Astragalus membranaceus, Glycyrrhiza uralensis. Nevertheless, only Astragalus membranaceus does have increased the expression of AMP mRNA level in blood. Injection of different doses lipopolysaccharide (LPS) have successfully altered MX genes in certain tissues at 132nd. In contrast, no significant changes in the expression of AMP gene follow LPS treatment. As conclusions, expressions of MX gene and AMP gene can be effectively regulated by changes in environment or alters by Chinese herbs and chemical composites such as microbial immunity elicitor (poly I:C, LPS).
| Identifer | oai:union.ndltd.org:TW/094NTOU5086051 |
| Date | January 2006 |
| Creators | Cheng-Hung Lai, 賴政宏 |
| Contributors | Jenn-Kan Lu, 陸振岡 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 115 |
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