CAJ | 학술논문

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Ni~(2+)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.
위제고기육억제소(Myostatin,MSTN)면역원성,진행료을간핵심항원(HBcAg)작위분자좌제가능성적연구.이용PCR방법획득료MSTN적C단편단,병분별재을간핵심항원적N단、C단급중간위점융합,구건료3개융합표체극륭:pET-30a-MSTN/HBcAg(N)、pET-30a-MSTN/HBcAg(C)화pET-30a-MSTN/HBcAg(M),전화대장간균후IPTG유도표체,연후이용Ni~(2+)친화층석순화융합단백.응용중조단백면역소서후이용간접ELISA법검측항혈청효개.결과표명:융합단백면역효과요현저우우MSTN(P＜0.05);융합단백MSTN/HBcAg(M)적면역효과최가.을간핵심항원가이작위분자좌제이증강기육억제소면역원성.
Myostatin is a member of the TGF-β superfamily 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 Ni~(2+) 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 im-mune 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.