碩士 / 國立海洋大學 / 水產養殖學系 / 88 / The increased prevalence of diseases has significantly reduced some important cultured shrimp production. Gene transfer techniques offer a powerful approach for changing the genetic traits of economic crustacean species for both basic research and commercial applications. Antimicrobial peptides are found in both myoloid cells and mucosal tissues of many vertebrates and invertebrates. These peptides are predicted to operate as a first-line host defense mechanism exerting broad-spectrum activity against pathogenic bacteria, fungi, parasites, and enveloped virus. Electroporation is a mass gene transfer method to transfer foreign genes into organisms. The objective of present study employs anti-microbial peptide (cecropin) protection aspects to kill and lyse bacteria to produce disease-resistant tiger shrimp (Penaeus monodon) by electroporation. The antimicrobial peptide mini gene constructs contained CMV (cytomegalovirus) promoter, catfish Ig M leader sequences, fused with the cecropin B cDNA and bGH poly A tail were used for production of transgenic shrimp. In present study, a BXT Electro Square Porator T820 petri dish electrodes model was used to transfer foreign DNA into newly fertilized eggs of tiger prawn. Optimal conditions for electroporation will be standardized using total 30,000~70,000 eggs placed into a petri dish with 10 mm electrode gap. 12 ml filtered sea-water containing linearized DNA fragments (480mg/ml) was used as the electroporation medium. The results show (a) cecropin B of artificial synthesized have bactericidal effects to E.coli, Vibrio harveyi and Vibrio alginolyticus. (b) hatching rate is effected by the strength of voltage. (c) 40-60% of the survived tiger shrimp carry transgene. (d) Transgene expression also has been demonstrated in PCR positive shrimp.
| Identifer | oai:union.ndltd.org:TW/088NTOU0086004 |
| Date | January 2000 |
| Creators | Tai-Hsin Yang, 楊泰鑫 |
| 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 | 0 |
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