人类组氨酰tRNA合成酶类似物的原核表达及其多克隆抗体的制备

目的：在原核系统中表达并纯化人类组氨酰tRNA合成酶类似物蛋白，制备多克隆抗体。 方法： 设计引物扩增其开放阅读框，重组入原核表达载体pQE30,并在大肠杆菌M15中诱导表达; 应用纯化的重组蛋白免疫新西兰大白兔。 结果： ①成功构建人类组氨酰tRNA合成酶类似物的原核表达载体；②经Ni亲和层析获得纯度较高的重组蛋白；③制备了高滴度的兔抗人类组氨酰tRNA合成酶类似物蛋白的多克隆抗体。 结论： 原核表达载体的构建、重组蛋白的表达、纯化及多克隆抗体的制备为今后研究该蛋白的功能提供了良好的基础。

Prokaryotic expression of the human histydyl-tRNA synthetase analogue and the preparation of its polyclonal antibody

AIM: To express and purify human histydyl - tRNA synthetase analogue and prepare its polyclonal antibody. METHODS: The open reading frame of the synthetase gene was amplified by PCR, then recombined into prokaryotic expression vector pQE30 and transformed into E. coli M15 for expression. Rabbits were immunized with purified expression products. RESULTS: Prokaryotic expression vector pQE30 of human histydyl - tRNA synthetase analogue was constructed. The recombined protein was purified by Ni affinity chromatography, and high titer polyclonal antibody was successfully prepared. CONCLUSION: Re-combined prokaryotic expression vector, the purified protein and prepared polyclonal antibody were the necessary materials for farther study of this protein.