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中国农学通报

中国农学通报 ›› 2008, Vol. 24 ›› Issue (10): 22-25.

所属专题： 生物技术； 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

鸡 SIgM λ 轻链蛋白在大肠杆菌中的表达与鉴定

赵绪永   

  • 收稿日期:2008-06-30 修回日期:1900-01-01 出版日期:2008-10-05 发布日期:2008-10-05

Expression and Identification of the expression of chicken surface IgM λ light chain in E.Coli

ZHAO Xu-Yong   

  • Received:2008-06-30 Revised:1900-01-01 Online:2008-10-05 Published:2008-10-05

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摘要： 应用 RT-PCR 技术从鸡法氏囊 B 细胞总 RNA 中克隆出 SIgM λ 轻链基因，并将其进一步克隆于 pET-28a 载体 T7 启动子的下游，构建了 pET-SIgM λ 原核表达载体。转化宿主菌 BL21(DE3)，经 IPTG 诱导，成功表达了25.5 kDa 的鸡 SIgM λ 轻链蛋白。经 SDS-PAGE 和 Western-blotting 分析， 表达产物以包涵体形式存在，且能与 His-Taq 单抗特异性反应，为进一步研究 SIgM 的免疫学功能及其他生物学功能奠定基础。

关键词: 杂交水稻, 杂交水稻, 博II优15, 选育, 应用

Abstract: The SIgM λ light chain gene about 630bp was obtained from the total RNA of the chicken bursal B lymphocyte by RT-PCR , then the SIgM λ light chain gene was subcloned into the downstream of T7 promoter of the vector pET-28a to construct expression vector of pET SIgM λ. The recombinant plasmid was transformed into in E. coli BL21 (DE3) and induced with IPTG. SDS-PAGE and Western blotting analyses showed that the chicken SIgM λ light chain protein of 25.5 kDa were efficiently expressed and mainly existed as inclusion bodies ,with specific reactivity to anti-His antibodies. Moreover，it may be helpful to study its Immunologic and other biological functions．