| 诱生性一氧化氮合酶cRNA探针的制备及敏感性检测 |
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| 引用本文： | 王迎伟,何球藻.诱生性一氧化氮合酶cRNA探针的制备及敏感性检测[J].徐州医学院学报,1999,19(4):265-267. |
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| 作者姓名： | 王迎伟 何球藻 |
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| 作者单位： | [1]徐州医学院免疫学教研室 [2]上海医科大学免疫学教研室 |
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| 基金项目： | 江苏省科委及卫生厅基金 |
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| 摘 要： | 目的 制备地高辛标记的大鼠诱生性一氧化氮合酶（ｉＮＯＳ）ｃＲＮＡ探针。方法 首先对携带大鼠ｉＮＯＳ ｃＤＮＡ的ＰＭＯＳ Ｂｌｕｅ质粒进行转化扩增。然后提取扩增的质粒ＤＮＡ并用Ｂａｍ ＨⅠ内切酶酶切处理使其线性化。以线性化的ｃＤＮＡ为模板，在Ｔ７ＲＮＡ聚合酶的作用下，体外转录带有地高辛的ｉＮＯＳ ｃＲＮＡ探针。
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| 关 键 词： | 地高辛 iNOS探针 质粒 一氧化氮合酶 cRNA |
THE PREPARATION AND DETECTION OF INDUCIBLE NITRIC OXIDE SYNTHASE (iNOS) cRNA PROBE |
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| Wang Yingwei,Gao Fengguang,He Qiuzao,et al.THE PREPARATION AND DETECTION OF INDUCIBLE NITRIC OXIDE SYNTHASE (iNOS) cRNA PROBE[J].Acta Academiae Medicinae Xuzhou,1999,19(4):265-267. |
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| Authors: | Wang Yingwei Gao Fengguang He Qiuzao |
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| Abstract: | Objective To prepare DIG labelled iNOS cRNA probe of rat. Methods First, the plasmid (PMOS Blue) carrying iNOS cDNA of rat was proliferated in E.coli HB101. Then, the DNA of the plasmid was isolated and linearized with Bam H restriction enzyme, Finally, the probe of iNOS cRNA was synthesized in vitro with T7 RNA polymerase in the presence of the template (the linearized cDNA) and DIG-UTP. Results The 0.1g/L (0.1pg/l) homologous cDNA was detected with the DIG labelled iNOS cRNA probe in dot-blot. Conclusion The iNOS cRNA probe thus prepared can be used for detecting the expression of iNOS mRNA in tissue cells. |
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| Keywords: | DigoxigeniniNOS probePlasmid |
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