碩士 / 輔仁大學 / 生物學研究所 / 86 / CDNAs encode mature grouper and ayu growth hormones were amplified by PCR from the grouper and ayu cDNA clones. The obtained PCR products were ligated into pET-20B(+) expression vector and expressed in E. coli, BL21(DE3) pLysS, under the T7 promoter regulation. Through IPTG induction, large amount of recombinant GH were produced and the size of the recombinant grouper and ayu GH are 194 and 195 amino acids, respectively. Since the expressed GH were packed in inclusion bodies, 0.4% Sarkosyl was used to dissolve the inclusion bodies and followed by dialysis to remove Sarkosyl. The recombinant peptides consist of 6 Histidines on the C-terminal for binding to His-tag binding resin. With this His-tag binding resin purification method, 10 mg/l recombinant GH peptides in more than 99% purity was obtained. In order to analyze the activity of recombinant GH, we injected tilapia intraperitoneally with 1μg 10μg per gram body weight of recombinant GH. We found that 10μg grouper recombinant GH induced higher level expression of IGF-I than 1μg grouper recombinant GH. However, injection of ayu recombinant GH into tilapia seems to have no promoting effect on the IGF-I expression level. Besides, we injected juveniles of grouper every week or every two weeks at concentration of 1μg and 10μg of these recombinant GH to each gram body weight of fish. We found that these two recombinant GH seem to have no growth promotion with weekly injection. However, with two weeks injection, 10 μg recombinant GH hormone promote fish growth very well.
| Identifer | oai:union.ndltd.org:TW/086FJU03112002 |
| Date | January 1998 |
| Creators | 蔡志東 |
| Contributors | , 陳擎霞, 張繼堯 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 76 |
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