| 番鸭白细胞介素2基因的克隆及其原核表达 |
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| 引用本文： | 吴异健,孟艳琴,吴宝成,黄一帆. 番鸭白细胞介素2基因的克隆及其原核表达[J]. 中国预防兽医学报, 2012, 34(2): 146-148 |
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| 作者姓名： | 吴异健 孟艳琴 吴宝成 黄一帆 |
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| 作者单位： | 1. 福建农林大学动物科学学院,福建福州,350002
2. 福建农林大学动物科学学院,福建福州350002;鄂尔多斯市农牧学校,内蒙古达拉特014300 |
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| 基金项目： | 福建省自然科学基金项目(Z0516012) |
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| 摘 要： | 为原核表达番鸭白细胞介素2 (MdIL-2),本研究利用ConA刺激诱导番鸭脾淋巴细胞,采用RT-PCR技术扩增MdIL-2基因的编码序列,并将其克隆至pGEX-6P-1中,构建重组质粒pGEX-MdIL-2,转化E.coli BL21受体菌.测序结果表明,MdIL-2基因ORF全长420 bp,编码140个氨基酸.重组菌经IPTG诱导表达后,SDS-PAGE分析显示,重组蛋白约为40 ku.Western blot分析表明,重组蛋白能够被抗GST单克隆抗体特异性识别.MdIL-2基因的克隆及其原核表达为进一步进行MdIL-2的活性检测以及应用研究奠定了基础.
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| 关 键 词： | 番鸭 白细胞介素2 克隆 表达 |
Cloning and expression of Muscovy duck interleukin-2 gene in Escherichia coli |
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| WU Yi-jian , MENG Yan-qin , WU Bao-cheng , HUANG Yi-fan. Cloning and expression of Muscovy duck interleukin-2 gene in Escherichia coli[J]. Chinese Journal of Preventive Veterinary Medicine, 2012, 34(2): 146-148 |
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| Authors: | WU Yi-jian MENG Yan-qin WU Bao-cheng HUANG Yi-fan |
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| Affiliation: | 1*(1.College of Animal Science,Fujian Agricultural and Forestry University,Fuzhou 350002,China; 2.Agricultural and Animal Husbandry School of Ordos,Dalate 014300,China) |
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| Abstract: | To express the Muscovy duck interleukin-2(MdIL-2) gene,the encoding sequence of IL-2 was amplified by RT-PCR from Muscovy duck spleen lymphocytes stimulated with concanavalin A,and the 420 bp ORF of MdIL-2 was inserted into pGEX-6P to constructed recombinant expression plasmid pGEX-MdIL-2 and expressed in E.coli by IPTG induction.SDS-PAGE analysis showed that the expressed product of the GST-MdIL-2 was 40 ku which was recognized by GST monoclonal antibody.The results could be used for further study of the bioactivity and application of MdIL2. |
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| Keywords: | muscovy duck interleukin-2 gene cloning expression |
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