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題 名 | Production of Recombinant Oreochromis Mossambicus Growth Hormone (GH) Polypeptides in E. Coli Cells and Characterization of the Molecular 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 |
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作 者 | 陳志毅; 吳金洌; | 書刊名 | 臺灣水產學會刊 |
卷 期 | 25:4 1998.12[民87.12] |
頁 次 | 頁251-263 |
分類號 | 388.5 |
關鍵詞 | 莫三鼻克; 生長激素; 基因體結構; 蛋白質表現; Oreochromis mossambicus; Growth hormone; Gene structure; Protein expression; |
語 文 | 英文(English) |
中文摘要 | 從吳郭魚(莫三鼻口孵魚)之腦下垂體抽取核醣核酸後,利用魚類生長激素專一 性極高之引子,當反轉錄時作用之引子,利用聚合�t連鎖反應選殖出莫三鼻口孵魚之生長激 素 cDNA,莫三鼻口孵魚之生長激素由 204 個胺基酸所構成,其核��酸序列與吳郭魚(尼羅 口孵魚)大約具有 98% 之相似性。pQE30 載體連接莫三鼻口孵魚之生長激素核��酸( cDNA )序列,利用大腸桿菌系統表現出大約 22kDa 之重組生長激素蛋白質。 莫三鼻口孵魚之基 因結構體由 6 個外顯子 (intron) 與 5 個內隱子 (exon) 所組成,與尼羅口孵魚之核��酸 序列相似性大約 97%。 |
英文摘要 | The complementary DNA (cDNA) encoding the prepeptide growth hormone (pre-GH) from tilapia (Oreochromis mossambicus; om) has been cloned and sequenced. The cDNA was derived by reverse transcription of RNA from total RNA of the pituitary gland using the method of polymerase chain reaction (PCR) and oligonucleotide primers specific for fish growth hormone as amplification primers. The omGH cDNA encoded a pre-hormone of 204 amino acid residues. The nucleotide sequence of omGH showed greater than 98% homology with that of Tilapia niloticus indicating a high degree of conservation for GH cDNAs sequence within the genus Tilapia. Mature omGH cDNA was inserted in an Escherichia coli expression vector (pQE30 vector, the pQE 30 vector containing a 6 × His affinity tag), and a protein of about 22 kDa was expressed in E. coli cells. The bacterially expressed omGH protein, contained within the inclusion body pellet, was dissolved in 8 M urea, and purified in a Ni-NTA column. Both the cloned tilapia (O. mossambicus) gene and that of T. niloticusis were divided into 6 exons and 5 introns. Comparison of the omGH genomic DNA sequence with that of Tilapia niloticus showed a homology of approximately 97%. This is significat from an evolutionary standpoint, suggesting a high degree of conservation for the GH gene within the genus Tilapia. |
本系統中英文摘要資訊取自各篇刊載內容。