中文摘要：

目的：本实验利用基因重组技术,构建融合Th细胞TT830-844表位序列的人重组Myostatin基因疫苗真核表达载体pVAC-TT-Ms,检测该基因疫苗对免疫小鼠前肢抓力的影响,为后续以Myostatin作为靶分子来改善废用性肌肉萎缩等病症的研究提供实验基础。方法：化学耦联方法合成Myostatin C-末端成熟肽（330 bp）基因序列及其N-端融合TT表位序列的DNA片段,重组质粒pVAC-TT-Ms的构建、纯化及瞬时转染中国仓鼠卵巢细胞（CHO细胞）按该类研究中的常规实验操作,并检测重组基因疫苗pVAC-TT-Ms免疫小鼠骨骼肌前肢抓力的变化。结果：酶切和测序结果表明,重组Myostatin基因疫苗表达载体pVAC-TT-Ms构建成功。重组质粒pVAC-TT-Ms OD260/280比值和电泳结果显示,该质粒作为基因疫苗符合免疫动物的质量要求。细胞荧光免疫检测结果表明,瞬时转染的CHO细胞中目的蛋白明显表达。动物免疫实验初步证明,重组Myostatin基因疫苗可显著提高免疫小鼠的前肢抓力,与正常对照组比较,免疫小鼠前肢抓力平均增加了29.88%。结论：本实验成功构建了在真核细胞中能有效表达融合TT表位序列人重组Myostatin基因疫苗表达载体,初步证明了重组Myostatin基因疫苗可显著增加免疫小鼠前肢抓力。

英文摘要：

Objective： To further study the therapy of wasting muscle by myostatin as a new targets,the eucaryotic expression vector coupled the foreign T-helper epitope of tetanus toxin（TT） to the N terminus of myostatin was constructed, and the effects of the gene vaccine on forelimb grip were tested in immunized mice.Methods： A DNA fragment encoding the TT epitope followed by the N terminus of mature myostatin（330bp） was synthesized.The eucaryotic expression vector of myostatin was constructed and the chinese hamster ovary（CHO） cells were infected with the recombinant plasmids pVAC-TT-Ms by liposome transfection according to routine laboratory procedure.The myostatin expression was tested by cell immunofluorescence technique in transfected CHO.The forelimbs grip were tested in immunized mice with myostatin gene vaccine.Results： The eucaryotic expression vector of myostatin coupled TT epitope was contructed successfully through the restriction analysis and sequencing.The recombinant plasmids pVAC-TT-Ms met quality criterion as gene vaccine by analysis OD260/280 and electrophoresis.The myostatin expression was detected obviously in transfected CHO.The forelimb grip in immunized mice had an obvious increase.The average value of forelimb grip of the mice immunized with pVAC-TT-Ms was about 29.88% greater than that of control mice.Conclusion： The construction of eucaryotic expression vector of myostatin coupled TT epitope is successful in expression for recombinant human mature peptide of myostatin.The gene vaccine of myostatin meet quality criterion.The immunized mice has an obvious increase in forelimb grip.