碩士 / 國立海洋大學 / 水產養殖學系 / 91 / Increased prevalence of infectious diseases has significantly reduced important cultured shrimp production. Gene transfer techniques offer a powerful approach for changing the genetic traits of economic animals for both basic research and commercial applications. Antimicrobial peptides were found in plants, vertebrates and invertebrates. These peptides were serve as a first-line host defense mechanism exerting broad-spectrum activity against pathogenic bacteria, fungi, parasites, and enveloped virus. The antisense RNA technique and coat protein protection assay were also been used to prevent the virus infection. In this study, transgenic constructs of Cecropin B, VP28-sense and VP28-antisense were used to produce transgenic penaeid shrimps. A new gene transfer method (in vivo spermatophore-mediated gene transfer; in vivo SMGT) was developed to generate disease resistant transgenic shrimps. Transgenic constructs were injected into the spermatophore, and then electroporation was employed to assist foreign DNA into oocytes. In vivo SMGT method was performed in the spermatophore of female tiger shrimps prior to spawning. On the other hand, in vivo SMGT method was performed in white shrimp spermatophore before mating. A BTX 2-Needle Array Electrode mode was used to transfer foreign DNA into the spermatophore of penaeid shrimps broods in this study. The results showed that the survival rate of in vivo SMGT method treated on female tiger shrimps (Penaeus monodon) broods and male white shrimps (Litopenaeus vannamei) broods were 100% and 85%, respectively. Hatching rate and survival rate of tiger shrimp larval (at postlarval PL6 stage) from in vivo SMGT method treated female tiger shrimps broods was 94% and 2%. Hatching rate and survival rate of white shrimp larval (at postlarval PL3 stage) produced by in vivo SMGT method were 85% and 50%. Genomic DNA PCR analysis indicated that thirty eight percent of surviving the transgenic tiger shrimps carried foreign genes. Ninety six percent of surviving white shrimps carried foreign genes. Expressing of cecropin B transgene was detected in 78% of transgenic white shrimps at PL28 stage.
| Identifer | oai:union.ndltd.org:TW/091NTOU0086002 |
| Date | January 2003 |
| Creators | Kao, Ching-Chi, 高靜琦 |
| Contributors | Lu, Jenn-Kan, 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 | 72 |
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