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Biochemical and immunological applications of a rat glutation transferase-histidine decarboxylase fusion protein

机译:大鼠凝集转移酶-组氨酸脱羧酶融合蛋白的生化和免疫学应用

摘要

Biochemical and immunological applications of a rat glutation transferase-histidine decarboxylase fusion protein. By reverse transcription (RT) and chain amplification (PCR) a rat histidine decarboxylase cDNA with residues from 1 to 512 (rHDC1/512) was obtained and this was used to construct a recombinant plasmid capable of expressing the fusion of glutation transferase (GST) with rHDC1/512. Purification of polypeptides with HDC activity is considerably simplified and cheapened through the use of this fusion (GST-rHDC1/512) in comparison with the acquisition of HDC protein from natural sources. The fusion protein can be used to search for and characterise antihistamine drugs which have histidine decarboxylase activity as a target and for the investigation of post- translational affinity, degradation and modification of the HDC polypeptide. Starting with the GST-rHDC1/512 fusion protein polyclonal antibodies capable of recognising the original rat HDC antigen were obtained, and these are of interest in the development of purification and diagnostic methods for the protein.
机译:大鼠凝集转移酶-组氨酸脱羧酶融合蛋白的生物化学和免疫学应用。通过逆转录(RT)和链扩增(PCR),获得了大鼠组氨酸脱羧酶cDNA,其残基为1至512(rHDC1 / 512),并将其用于构建能够表达谷氨酸转移酶(GST)融合体的重组质粒。使用rHDC1 / 512。与从天然来源获得HDC蛋白相比,通过使用这种融合蛋白(GST-rHDC1 / 512)大大简化了具有HDC活性的多肽的纯化并降低了成本。该融合蛋白可用于搜索和表征具有组氨酸脱羧酶活性作为靶标的抗组胺药,以及用于研究翻译后亲和力,HDC多肽的降解和修饰。从GST-rHDC1 / 512融合蛋白开始,获得了能够识别原始大鼠HDC抗原的多克隆抗体,这些对于开发纯化和诊断该蛋白的方法具有重要意义。

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