雞介白素-2之重組蛋白表現與多株抗體製備

Recombinant Protein Expression and Polyclonal Antibody Development of Chicken Interleukin-2

蔡淑婷(Shu-Ting Tsai) ； 范怡馨(Yi-Hsin Fan) ； 李龍湖(Long-Huw Lee) ； 邱垂章(Chwei-Jang Chiou) ； 謝快樂(Happy K. Shieh) ； 邱繡河(Shiow-Her Chiou)

《臺灣獸醫學雜誌》 31卷1期 (2005/03) Pp. 38-45

https://doi.org/10.7009/TVJ.200503.0038

摘要

研究目的是表現雞介白素-2（chicken interleukin-2, chlL2）之重組蛋白以及製備其多株抗體。我們以大豆素A（concanavalin A, ConA）刺激雞的周邊血白血球， 萃取RNA，進行反轉錄聚合臨鏈反應（reverse transcription-polymerase chain reaction, RT-PCR）選殖chlL2之cDNA基因，構築chlL2原核表現貿體pETchlL2，送入大腸桿菌誘導重組的chlL2（recombinant chlL2, rchlL2）蛋白之表現，純化之重組蛋白由質譜儀分析確認為rchlL2。其次，將rchlL22蛋白施打於小鼠，產生的多株抗體以西方墨潰法分析證實可辨識的chlL2蛋白。本研究己成功表現rchlL2蛋白及製備其多株抗體，將來可提供於獸醫防疫或相關研究。

關鍵字

雞介白素-2 ； 細胞素 ； 多株抗體 ； 雞介白素-2重組蛋白

並列摘要

The goals of this study are to express recombinant protein of chicken interleukin-2 (chlL2) and to develop polyclonal antibody specific for chlL2. We stimulated chicken peripheral leukocytes with concanavalin A (ConA), extracted RNA, and employed reverse transcription-polymerase chain reaction (RT-PCR) to clone cDNA of chlL2. A prokaryotic expression plasmid of chlL2, designated as pETch1L2, was constructed, introduced into Escherichia coli to induce recombinant chlL2 (rchlL2) protein expression. Purified rchlL2 was further analyzed by mass spectrometry and its identity was confirmed. Moreover, we immunized mice with rchlL2 to elicit anti-chlL2 polyclonal antibody. Western blot analysis showed that the resultant mouse polyclonal antibody recognized chlL2 with specificity. In conclusion, in this study, we have developed rchlL2 protein and anti-chlL2 polyclonal antibody. Both reagents can be provided for disease control in veterinary medicine or related research. The goals of this study are to express recombinant protein of chicken interleukin-2 (chlL2) and to develop polyclonal antibody specific for chlL2. We stimulated chicken peripheral leukocytes with concanavalin A (ConA), extracted RNA, and employed reverse transcription-polymerase chain reaction (RT-PCR) to clone cDNA of chlL2. A prokaryotic expression plasmid of chlL2, designated as pETch1L2, was constructed, introduced into Escherichia coli to induce recombinant chlL2 (rchlL2) protein expression. Purified rchlL2 was further analyzed by mass spectrometry and its identity was confirmed. Moreover, we immunized mice with rchlL2 to elicit anti-chlL2 polyclonal antibody. Western blot analysis showed that the resultant mouse polyclonal antibody recognized chlL2 with specificity. In conclusion, in this study, we have developed rchlL2 protein and anti-chlL2 polyclonal antibody. Both reagents can be provided for disease control in veterinary medicine or related research.

並列關鍵字

chicken inter/eukin-2 (chlL2) ； cytokine ； polyc/onal antibody ； recombinant chlL2 protein (rchlL2 protein)