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目的：获取旋毛虫ＥＳ抗原的结构基因，对其鉴定和克隆，并研制旋毛虫基因重组抗原。方法：先用反转录ＰＣＲ技术获取目的基因，经序列测定和酶切分析后，再用重组ＤＮＡ技术分别将目的基因与融合表达载体ｐＥＸ３１Ｃ、ｐＥＸ３１Ｂ及表达载体ｐＢＶ２２０连接，评价在大肠杆菌中的表达效果和鉴定表达产物的特异性。结果：获得了编码ＥＳ抗原特异性蛋白成分的两个结构基因（０．７ｋｂ和０．９５ｋｂ），其序列与文献报道的稍有差异。共构建３个重组质粒并在大肠杆菌中表达出相应分子量大小的重组蛋白，均能被猪旋毛虫病阳性血清所识别，但非融合蛋白的特异性强于融合蛋白。在相同条件下，融合蛋白的表达量高于非融合蛋白，而表达蛋白的分子量大小与表达水平呈负相关性。结论：在大肠杆菌中表达的３种重组蛋白是研制旋毛虫基因重组抗原的良好候选抗原蛋白。
OBJECTIVE: To obtain the structural gene of Trichinella ES antigen, identify and clone it, and develop recombinant Trichinella antigen. Methods: The target gene was obtained by reverse transcription-polymerase chain reaction (RT-PCR), and sequenced and digested with restriction endonuclease. The recombinant plasmid was used to ligate the target gene with pEX31C, pEX31B and pBV220 respectively. Expression effect and identification of the specificity of the expression product. RESULTS: Two structural genes (0.7 kb and 0.95 kb) encoding ES antigen-specific protein components were obtained, and their sequences were slightly different from those reported in the literature. A total of three recombinant plasmids were constructed and expressed in Escherichia coli. The corresponding molecular weight recombinant proteins were all recognized by Trichinella spiralis positive sera, but the specificity of the non-fusion protein was stronger than that of the fusion protein. Under the same conditions, the expression level of the fusion protein was higher than that of the non-fusion protein, and the molecular weight of the expressed protein was negatively correlated with the expression level. CONCLUSION: The three recombinant proteins expressed in E. coli are good candidate antigenic proteins for the development of recombinant Trichinella antigens.