Transgenesis of creatine kinase using cold-inducible and muscle-specific system to develop cold tolerance in zebrafish

• Main Authors: Ta-Huei Lin, 林大暉
• Other Authors: Jen-Leih Wu
• Format: Others
• Language: en_US
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/79181847604785000341

碩士 === 國立臺灣師範大學 === 生命科學研究所 === 94 === The fish that lives in the tropical or subtropical area is adapted in the temperature of the warm environment and damaged by the low temperature in the winter. In order to confront the economic loss caused by the cold stress, the improvement of the cold tolerance of fish has been studied. We analyzed the biochemical function of the M3-CK sub-isozyme from carp (Cyprinus carpio, a eurythermic fish) and found out the M3-CK was stable and active in low temperature. The M3-CK is the key enzyme of energy metabolism of carp in low temperature. We constructed a dual functional expression vector that constitutively expressed the carp MCK and a GFP marker gene. The CMV promoter drove the GFP marker and the CMV promoter, cold-inducible promoter and carp M3-CK promoter drove the carp MCK isozymes, M1-CK and M3-CK, respectively. By transgenesis of the key enzyme, carps M3-CK, into the model fish, zebrafish (Danio rerio), we found that the transgenic fish can express M3-CK constitutively and enhance its cold tolerance from demonstrating a substantial improvement in swimming ability in 13℃. But transgenesis of the carps M1-CK into the model fish zebrafish (Danio rerio); we found that the transgenic fish don’t improve its cold tolerance in swimming ability in 13℃. It is a proper target to help the tropical or subtropical fish confront the cold stress by maintaining the metabolic capability in low temperature. The zebrafish is a good model system for the transgenesis of the enzyme to improve the cold tolerance of tropical and subtropical fishes. Statistically, the results in Table 7 revealed significant differences in the swimming velocity between the wild type and the transgenic zebrafish at the lower temperature. The cold-inducible promoter and M3CK promoter can synthesis carp M3-CK that increase swimming ability of transgenic zebrafish at the right moment. The results show that it is work for the assumption that M3-CK is able to improve the target fish’s cold tolerance ability. Consequently, this application of the zebrafish is a clue for the application of economic species of fish to confront the low temperature.