Production of Recombinant Oreochromis Mossambicus Growth Hormone(GH)Polypeptides in E. Coli Cells and Characterization of the Moloecular Structure of the GH Gene

吳郭魚（莫三鼻口孵魚）生長激素之基因選殖，蛋白質表現與基因結構體定序分析

陳志毅(Jyh-Yih Chen) ； 吳金洌(Jen-Leih Wu)

《臺灣水產學會刊》 25卷4期 (1998/12) Pp. 251-263

https://doi.org/10.29822/JFST.199812.0002

摘要

吳郭魚精液先以精虫抑制活化緩衝液清洗處理後，再懸浮於0.85％淡水魚生理食鹽水中，用於電穿孔處理時做為轉殖基因的接受者。這些電穿孔處理過帶有轉殖基因的精子，立刻以液態氮冷凍保存達40天。解凍後的精子與成熟吳郭魚卵授精，解凍後的精子其孵化率為23.79％較用新鮮的精子授精的孵化率43.25％為差。解凍後的精子細胞，內含轉殖基因可利用南方氏核苷酸雜交法檢測到。又授精後孵化的小魚，於萃取核苷酸做聚合酶連鎖反應法檢測，亦可証實轉殖基因的存在，因此冷凍保存電穿孔處理過帶有轉殖基因的精子，可做為轉殖基因的攜带者直接用於人工授精製作基因轉殖吳郭魚。

關鍵字

莫三鼻克 ； 生長激素 ； 基因體結構 ； 蛋白質表現

並列摘要

Tilapia milt treated with sperm deactivator and suspended in 0.85% freshwater fishsaline was used as the DNA recipient for gene transfer by electroporation.These electro-porated sperm were immediately cryopreserved in liquid nitrogen and stored for 40 days.As measured by hatching percentages,the viability of thawed cryopreserved sperm usedfor fertilization was 23.79% less than that 43.25% of fresh sperm.The presence of thetransgene within the post-thawed electroporated sperm was confirmed by Southern blot,and the presence of the transgene in the genome of transgenic fry was confirmed by PCR.We conclude that cryopreserved electroporated tilapia sperm is a good transgene mediatorthat can be used to generate transgenic tilapia.

並列關鍵字

Oreochromis Mossambicus ； Growth Hormone ； Gene Structure ； Protein Expres-sion