兔岩藻糖基转移酶基因的克隆与原核表达

【目的】克隆兔岩藻糖基转移酶(FUT2)基因，并进行原核表达，为进一步研究RHDV的感染过程和致病机理奠定物质基础。【方法】从兔肌肉组织中抽提基因组DNA作为模板，PCR扩增出FUT2基因，然后将该基因亚克隆入原核表达载体pET30a中，获得重组原核表达质粒pET30a-FUT2。将该重组原核表达质粒转化E.coli BL21(DE3)感受态细胞，用IPTG进行诱导表达，分别用SDS-PAGE和Western-blot方法对表达产物进行鉴定。【结果】成功克隆了FUT2基因，该基因长度为1 044 bp。成功构建了原核重组表达质粒pET30a-FUT2，其诱导表达产物分子质量约为45 ku；表达的重组FUT2蛋白可与His标签抗体发生抗原抗体反应。【结论】成功克隆了兔岩藻糖基转移酶基因，获得了分子质量约为45 ku的FUT2蛋白。

Cloning and prokaryotic expression of rabbit fucosyltransferases

【Objective】 The research cloned and expressed Fucosyltransferases(FUT2),to lay the basis for further exploring the infection process and pathogenesis mechanism of Rabbit hemorrhagic disease virus.【Method】 Extraction genome from rabbit muscle tissue,FUT2 gene was synthesized and sub-cloned into the pET30a vector,and then the recombinant plasmid pET30a-FUT2 transformed into E.coli.BL21(DE3).The expression of the recombinant plasmid was induced by IPTG and analyzed by SDS-PAGE and Western-blot.【Result】 The results show that FUT2 gene is 1 044 bp in length,and the molecular weight of the recombinant protein,fused to his tag,is about 45 ku.【Conclusion】 FUT2 gene is successfully cloned,and the molecular weight of the recombinant protein is about 45 ku.