应用乙肝核心抗原增强肌肉抑制素免疫原性的研究

doi:10.7668/hbnxb.2009.05.041

华北农学报 ›› 2009, Vol. 24 ›› Issue (5): 197-200. doi: 10.7668/hbnxb.2009.05.041

应用乙肝核心抗原增强肌肉抑制素免疫原性的研究

马毅¹, 陈小强², 王文杰¹, 李晴³, 关宏³, 安晓荣³, 陈永福³

1. 天津市畜牧兽医研究所, 天津 300112;
2. 天津农学院, 天津 300384;
3. 中国农业大学, 农业生物技术国家重点实验室, 北京 100094

收稿日期:2009-04-22 出版日期:2009-10-28
作者简介:马毅(1976-)，男，天津人，博士，副研究员，主要从事动物基因工程研究。
基金资助:
天津市自然科学基金(08JCYBJC04500)

Study on the Improvement of Immunogenicity of Myostatin with Hepatitis B Core Antigen

MA Yi¹, CHEN Xiao-qiang², WANG Wen-jie¹, LI Qing³, GUAN Hong³, AN Xiaorong³, CHEN Yong-fu³

1. Tianjin Institute of Animal Science and Veterinary, Tianjin 300112, China;
2. Tianjin Agricultural University, Tianjin 300384, China;
3. State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, China

Received:2009-04-22 Published:2009-10-28

摘要/Abstract

摘要： 为提高肌肉抑制素(Myostatin，MSTN)免疫原性，进行了乙肝核心抗原(HBcAg)作为分子佐剂可能性的研究。利用PCR方法获得了MSTN的C端片段，并分别在乙肝核心抗原的N端、C端及中间位点融合，构建了3个融合表达克隆：pET-30a-MSTN/HBcAg(N)、pET-30a-MSTN/HBcAg(C)和pET-30a-MSTN/HBcAg(M)，转化大肠杆菌后IPTG诱导表达，然后利用Ni²⁺亲和层析纯化融合蛋白。应用重组蛋白免疫小鼠后利用间接ELISA法检测抗血清效价。结果表明：融合蛋白免疫效果要显著优于MSTN(P＜0.05)；融合蛋白MSTN/HBcAg(M)的免疫效果最佳。乙肝核心抗原可以作为分子佐剂以增强肌肉抑制素免疫原性。

关键词: 肌肉抑制素, 乙肝核心抗原, 融合蛋白, 免疫原性

Abstract: Myostatin is a member of the TGF-βsuperfamily that functions as a negative regulator of skeletal muscle development in mammals.Targeting the myostatin pathway may be an effective strategy for increasing muscle growth.To improve immunogenicity of myostatin,the possibility of hepatitis B core antigen as molecular adjuvant was studied.First,myostatin C-domain was amplified by PCR and fused to the gene of hepatitis B core antigen at the position of N-terminal,C-terminal and internal respectively.Then three expression vector pET-30a-MSTN/HBcAg (N),pET-30a-MSTN/HBcAg(C) and pET-30a-MSTN/HBcAg(M)was constructed.The fused proteins were expressed in E.coli. and were purified by Ni²⁺ affinity chromatography.Then male Kunming white mice were immunized with single fused protein or recombinant C-domain.The indirect ELISA assay was used to detect the titer of antiserum against myostatin.The results shown:The immune effect of fused proteins was significantly better than that of recombinant C-domain(P<0.05);The immune effect of fused protein MSTN/HBcAg(M)was the best in three fused protein.These results proved that HBcAg could be used as molecular adjuvant to improve the immunogenicity of myostatin.

Key words: Myostatin, HBcAg, Fused protein, Immunogenicity

中图分类号:

S852.4+4

马毅, 陈小强, 王文杰, 李晴, 关宏, 安晓荣, 陈永福. 应用乙肝核心抗原增强肌肉抑制素免疫原性的研究[J]. 华北农学报, 2009, 24(5): 197-200. doi: 10.7668/hbnxb.2009.05.041.

MA Yi, CHEN Xiao-qiang, WANG Wen-jie, LI Qing, GUAN Hong, AN Xiaorong, CHEN Yong-fu. Study on the Improvement of Immunogenicity of Myostatin with Hepatitis B Core Antigen[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(5): 197-200. doi: 10.7668/hbnxb.2009.05.041.

http://www.hbnxb.net/CN/Y2009/V24/I5/197

参考文献

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