豬上皮細胞生長因子之基因選殖、表現與生物活性

Gene Cloning, Expression, and Bioactivity of Porcine Epidermal Growth Factor

郭村勇(Tsun-Yung Kuo) ； 吳忠晉(Jhong-Jin Wu) ； 李德南(Der-Nan Lee) ； 吳輔祐(Fu-Yu Wu)

《宜蘭大學生物資源學刊》 2卷1期 (2005/12) Pp. 37-45

https://doi.org/10.6175/job.2005.21.15

摘要

上皮細胞生長因子（EGF）是人乳中的主要生長因子，有助幼小動物建立胃腸黏膜細胞屏障，也許可用於改善仔豬離乳下痢問題，有很大的經濟潛力。本實驗抽取豬顎下唾腺mRNA，以RT-PCR增幅得預期180 bp大小的基因片段，經基因定序確定與已發表的pEGF基因完全相同。此基因與pET24a質體重組後轉型至BL-21大腸桿菌，以IPTG誘導表現，經分離、溶解、透析可得相當產值之復性pEGF蛋白質。經測試其影響細胞生長狀況，證實有促進細胞生長的活性。

關鍵字

豬上皮細胞生長因子 ； 基因選殖 ； 表現 ； 生物活性

並列摘要

Epidermal growth factor (EFG) is the major growth factor in human milk. EGF may help to establish the gastrointestinal mucous barrier, which may improve the weaning pig diarrhea problem, thus possess economic profits. Pig submaxillary gland was collected for mRNA extraction. RT-PCT was conducted to amplify the expected pEGF gene with the result of an 180bp fragment. Subsequent gene sequencing showed the fragment is identical to the published pEGF gene. The cloned gene was inserted into pET24a plasmid and was transformed into BL-21 E. coli. Induction of gene expression by IPTG resulted in the production of EGF accumulated in inclusion body. Isolation, dispersion, and dialysis of the inclusion body produced abundant pEGF protein. With cell culture essay, the pEGF protein was demonstrated to have growth factor activity.

並列關鍵字

pEGF ； gene cloning ； expression ； bioactivity