| 鸡CD4基因片段原核表达及其单抗制备 |
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| 引用本文： | 程航,余为一. 鸡CD4基因片段原核表达及其单抗制备[J]. 中国家禽, 2011, 33(8) |
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| 作者姓名： | 程航 余为一 |
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| 作者单位： | 安徽农业大学安徽省人兽共患病重点实验室,安徽合肥,230036 |
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| 摘 要： | 将PCR扩增的鸡CD4基因片段克隆到pET-32a原核表达载体,构建获得原核表达重组载体pET-32a-CD4.重组载体导入大肠杆菌BL21(DE3),经过IPTG诱导表达,Ni-NTA亲和树脂纯化获得融合蛋白His-CD4.以此融合蛋白为抗原免疫BALB/c小鼠,用杂交瘤细胞技术制备了1株能够稳定传代并分泌抗CD4多肽单抗的杂交瘤细胞株,命名为3E12.经检测该抗体亚类为IgG2b,单抗腹水的间接ELISA效价为1:105,Western blot结果显示单抗与CD4多肽能特异性地结合.
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| 关 键 词： | 鸡 原核表达 单抗 |
Prokaryotic Expression of Gene Fragment of Chicken CD4 and Preparation of Monoclonal Antibody Against the Protein |
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| CHENG Hang,YU Weiyi. Prokaryotic Expression of Gene Fragment of Chicken CD4 and Preparation of Monoclonal Antibody Against the Protein[J]. China Poultry, 2011, 33(8) |
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| Authors: | CHENG Hang YU Weiyi |
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| Affiliation: | CHENG Hang,YU Weiyi**(Key Laboratory for Zoonoses in Anhui Province,Anhui Agricultural University,Hefei,Anhui 230036) |
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| Abstract: | The gene fragment of chicken CD4 was obtained by PCR and was cloned into pET-32a to construct recombinant prokaryotic expression vector pET-32a-CD4.And then the recombinant vector was transformed into Escherichia coli BL21(DE3).Fusion protein His-CD4 was induced by IPTG and purified with affinity Ni-charged resin.A hybridoma cell line secreting monoclonal antibody(McAb) against polypeptide CD4 was produced by fusing mouse myeloma cells(SP2/0) with spleen cells from BALB/c mice immunized with the recombinant... |
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| Keywords: | CD4 |
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