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首页> 外文期刊>Chemical Research in Chinese Universities >Cloning and Expression of Intracellular Part of Receptor Protein Tyrosine Phosphatase RPTPα and Preparation of Its Polyclonal Antibodies
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Cloning and Expression of Intracellular Part of Receptor Protein Tyrosine Phosphatase RPTPα and Preparation of Its Polyclonal Antibodies

机译:酪氨酸磷酸酶RPTPα胞内部分的克隆表达及多克隆抗体的制备

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摘要

A DNA fragment encoding the intracellular part of tyrosine phosphatase RPTPα designated as RPTPα-2D gene was amplified by PCR from a human prostate cDNA library and cloned into the pT7 E. coli expression vector. The resulting plasmid pT7-RPTPα-2D was used to transform Rosetta DE3 E. coli cells. RPTPα-2D was predominately expressed in the insoluble inclusion body and was effectively purified using preparative electrophoresis gels. Polyclonal antibodies were obtained after immunization of a rabbit with purified RPTPα-2D. The antibodies displayed a high titer and sensitivity. This study thus provided a valuable tool for further researches on RPTPα.
机译:通过PCR从人前列腺cDNA文库中扩增出编码酪氨酸磷酸酶RPTPα的称为RPTPα-2D基因的细胞内部分的DNA片段,并将其克隆到pT7大肠杆菌表达载体中。所得质粒pT7-RPTPα-2D用于转化Rosetta DE3大肠杆菌细胞。 RPTPα-2D主要在不溶性包涵体中表达,并使用制备性电泳凝胶进行了有效纯化。用纯化的RPTPα-2D免疫兔子后获得多克隆抗体。抗体显示出高滴度和敏感性。因此,这项研究为进一步研究RPTPα提供了有价值的工具。

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