http://scholars.ntou.edu.tw/handle/123456789/14440
標題: | Introducing foreign DNA into tiger shrimp (Penaeus monodon) by electroporation | 作者: | Tseng, F.S. Tsai, H.J. I-Chiu Liao Song, Y.L. |
公開日期: | 十二月-2000 | 出版社: | Theriogenology | 起(迄)頁: | 54, pp.1421-1432 | 摘要: | Electroporation was used to introduce pFLAG-CMV-1-BAP, a DNA fragment that includes a bacterial alkaline phosphatase gene driven by a human cytomegalovirus (CMV) promoter, into Penaeus monodon zygotes. The transgenic tiger shrimp was achievedby using 10kV, 28 pulses, 120 g sec pulse time, 10 cycles, and a DNA concentration of 37.5 microg/mL. The hatching rate of electroporated zygotes (46%) was significantly lower than that of zygotes in the untreated group (89%). The survival rate of postlarvae in the electroporated group using a DNA concentration of 37.5 microg/mL decreased from 0.6% for postlarva 45 to 0.4% for postlarva 120. Based on dot blot analysis, the rate of gene transfer was 37% in mysis-stage, 23% postlarva 15(PL15), 19% postlarva 45(PL45), and 21% 4-month-old (about PL120). Genomic Southern blotting demonstrated that DNA from transgenic tiger shrimp contained fragments of exogenous DNA that were smaller, larger and of the same molecular size as pFLAG-CMV-1-BAP. Transferred DNA fragments were integrated into the genomes of 31% of the transgenic tiger shrimp. The exogenous DNA was mosaically distributed in a wide variety of tissues. Immunohistochemical staining revealed that the FLAG-BAP fused-protein encoded by pFLAG-CMV-1-BAP was present in the ovaries of some transgenic tiger shrimp. |
URI: | http://ntour.ntou.edu.tw:8080/ir/handle/987654321/45043 http://scholars.ntou.edu.tw/handle/123456789/14440 |
DOI: | 10.1016/s0093-691x(00)00464-7 |
顯示於: | 海洋中心 |
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