日本血吸虫脂肪酸结合蛋白基因在大肠杆菌的表达

GENE EXPRESSION OF FATTY ACID-BINDING PROTEIN FROM SCHISTOSOMA JAPONICUM IN ESCHERICHIA COLI

目的 克隆并表达日本血吸虫脂肪酸结合蛋白基因以制备血吸虫疫苗候选分子。方法 利用 Ｐ Ｃ Ｒ 技术扩增日本血吸虫大陆株的ｃ Ｄ Ｎ Ａ 基因， 经 Ｂａｍ Ｈ Ｉ和 Ｅｃｏ Ｒ Ｉ双酶切后定向克隆于原核表达质粒 Ｐ Ｅ Ｇ Ｘ２ － Ｔ， 并在大肠杆菌进行表达研究。结果 获得目的基因克隆， 在大肠杆菌可诱导表达约４４ｋ Ｄ 的融合蛋白， 该重组抗原能被慢性血吸虫病人血清特异性识别。结论 成功地表达了一种血吸虫疫苗候选抗原， 为进一步研究打下基础。

Aim\ Cloning and expression the fatty acid-binding protein (FABP) gene of schistosoma japonicum to produce a candidate vaccine antigen for schitosomiasis.Methods\ the cDNA gene of S japonicum was amplified by a pair of primers designed according to the FABP gene,and the PCR products was digested with restrition endonuclease BamHI and EcoRI respectively for a directional gene cloning to the expression vector PEGX2-T,then studying its expression in E coli.Results\ The objective gene was successfully cloned and the sequence analysis of this gene revealed 99 3% identity with the reported sequence.It could be induced to express a 44kD fusion protein in E.coli and the recombinant antigen could be recognized differentially by the sera from chronic schitosomiasis patient.Conclution\ A schistosomiases condidate vaccine molecular has been expressed and it has laid the foundation of further research.