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Arah1是一种丰富的花生蛋白质 ,也是花生的一种主要过敏原。本实验从花生中提取totalRNA ,通过逆转录合成cDNA ,并根据文献报告的Arah1蛋白的基因序列合成两端引物 ,通过PCR方法进行基因的扩增 ,随后将其克隆到原核表达载体pRSET B上 ,在大肠杆菌中进行了表达。重组过敏原rArah1基因的序列分析结果表明 ,所获得的基因与文献资料相比是吻合的。免疫印迹实验表明重组的Arah1蛋白具有与花生过敏病人血清池中的IgE结合的活性
Arah1 is a rich peanut protein that is also a major peanut allergen. In this experiment, totalRNA was extracted from peanut, cDNA was synthesized by reverse transcription, and the double-stranded primer was synthesized according to the reported sequence of Arah1 protein. The gene was amplified by PCR and cloned into prokaryotic expression vector pRSETB. Expressed in E. coli. Sequence analysis of recombinant allergen rArah1 gene showed that the obtained genes are in good agreement with the literature data. Western blotting experiments showed that the recombinant Arah1 protein has IgE-binding activity in the serum pool of peanut-allergic patients