傳染性支氣管炎病毒S1基因之表現

The Expression of Infectious Bronchitis Virus S1 Gene

翁敏召(Min-Chao Weng) ； 王金和(Ching-Ho Wang)

《中華民國獸醫學會雜誌》 25卷1期 (1999/03) Pp. 34-42

https://doi.org/10.7009/JTCSVS.199903.0034

摘要

為發展雞傳染性支氣管炎病毒次單位疫苗，本實驗嘗試進行該病毒之S1醣蛋白基因的表現，奠定發展的基礎。依據已定序完成的臺灣分離株S1醣蛋白基因之核甘酸序列，設計二組引子，包含抗原決定位及較保留的區域。首先以這些引子進行RT-PCR反應，增幅出片斷大小為539 bp及331 bp的產物，將此產物與載體pRSET A, B, C和pGEX-4T-2進行接合作用，並經轉化作用至勝任細胞（大腸桿菌）內，再利用IPTG誘導細胞內質體表現S1基因的蛋白質，結果兩段基因成功的表現出融和蛋白質，分子量分別約為27.5 kDa和43 kDa的產物。

關鍵字

傳染性支氣管炎 ； S1基因 ； 表現

並列摘要

In order to develop a subunit vaccine of infectious bronchitis virus, parts of S1 glycoprotein were expressed in this experiment. Two sets of primers were designed according to the entire nucleotide sequence of SI glycoprotein of Taiwan isolate. Products included both hy-pervariable and conserved regions. RT-PCR products with 539 bp and 331 bp were amplified using these primers. The fragments were ligated in pRSET A, 8, C or pGEX-4T-2 vectors. Plasmids were introduced into competent cell (”E. coli”) by transformation, and the cells were induced by utilizing IPTG for the protein expression of SI gene. As a result, fusion proteins of two fragments were expressed successfully. Molecular weights of products were 27.5 kDa and 43 kDa, respectively.

並列關鍵字

Infectious bronchitis ； S1 gene ； Expression