中国农业科学 ›› 2007, Vol. 40 ›› Issue (1): 190-195 .

• 畜牧·兽医·资源昆虫 • 上一篇    下一篇

鹅γ-干扰素成熟蛋白的表达及其生物学活性研究

李洪涛,马波,王君伟,金红岩   

  1. 东北农业大学动物医学院
  • 收稿日期:2005-09-19 修回日期:1900-01-01 出版日期:2007-01-10 发布日期:2007-01-10
  • 通讯作者: 王君伟

Expression of goose interferon γ mature protein and detection of recombinant proteins bioactivity

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  1. 东北农业大学动物医学院
  • Received:2005-09-19 Revised:1900-01-01 Online:2007-01-10 Published:2007-01-10

摘要: 【目的】检测重组鹅IFN-γ的生物学活性。【方法】将鹅IFN-γ成熟蛋白基因分别克隆到原核表达载体pET30a,杆状病毒转移载体pMelBacA和pBlueBacHis2A。对鹅IFN-γ成熟蛋白进行原核和真核表达系统表达;利用体外活性试验检测表达产物的生物学活性。【结果】原核和真核表达系统表达了重组鹅IFN-γ成熟蛋白,并制备其原核表达产物的多克隆抗体;重组鹅成熟IFN-γ可以诱导鹅巨噬细胞产生NO,并能抑制GPMV在鹅胚成纤维细胞中的增殖。【结论】重组鹅成熟IFN-γ具有鹅巨噬细胞激活活性和抗病毒活性。

关键词: 干扰素(IFN), 成熟蛋白, 原核表达, 真核表达, 生物学活性

Abstract: the IFN genes, which encode goose interferon γ mature protein, were cloned from total RNA was derived from PHA-stimulated-cultured (PBMCs) of goose by reverse transcription polymerrase chain reaction (RT-PCR). The amplified products of PCR were cloned into pMD18-T vector and sequenced. Compared with the published goose IFN-γ genes sequence, the homology of nucleotide sequence were 100.00%. The fragments of IFN-γ were subcloned into plasmid pET30a and transfer plasmid pMelBacA, pBlueBacHis2A, respectively. the purified recombinant baculovirus were obtained. goose interferon γ mature proteins were successfully expressed in the prokaryotic and eukaryotic expression systems, respectively. And prepared polyclonal antibodies against the products of the prokaryotic expression. There have the common antigenic between the products of prokaryotic and eukaryotic expression systems. One of the biological activity of recombinant mature goose IFN-γ is the ability of activating macrophage of goose to generate nitric oxide(NO) was detected in vitro.

Key words: Interferon(IFN), mature protein, prokaryotic expression, eukaryotic expression, biological activity