大鼠组织中三磷酸肌醇受体亚型基因的表达
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Expression of Inositol Trisphosphate Receptor Subtypes mRNA in Rat Brain, Myocardium and Cultured Vascular Smooth Muscle Cells
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    摘要:

    为观察大鼠心肌、小脑和培养的主动脉血管平滑肌细胞中三磷酸肌醇受体亚型的基因表达,用异硫氰酸胍- 酸酚氯仿一步法抽提组织总R N A,用特异性引物进行逆转录- 聚合酶链式反应,β- actin 为内标半定量检测三磷酸肌醇受体亚型m R N A 的表达。将I I型P C R 产物重组、克隆并测序。发现三磷酸肌醇受体三个亚型在心肌、小脑和平滑肌细胞中均有表达,小脑组织中表达水平较高。P C R 扩增片段分别为: Ⅰ型三磷酸肌醇受体525 bp ( 小脑) 或405 bp ( 心肌和平滑肌细胞) ; Ⅱ三磷酸肌醇受体405 bp ;Ⅲ型三磷酸肌醇受体169 bp 。序列分析发现克隆的Ⅱ三磷酸肌醇受体c D N A 序列与设计的目的c D N A一致,提示建立的逆转录- 聚合酶链式反应方法能可靠地研究动物组织中三磷酸肌醇受体不同亚型的基因表达。

    Abstract:

    Aim To establish a RT-PCR method for detecting expression of mRNA for inositol trisphosphate receptor (IP 3R) and to investigate expression of IP 3R subtypes in rat brain, myocardium and cultured vascular smooth muscle cells (VSMC). Methods Total RNA was isolated from different tissue of rat and was reversibly transcripted into cDNA. RT-PCR was performed with β-actin as internal label. Three oligonuclotide primers for each subtype of IP 3R were employed to amplify predicted DNA. Results It was found that all subtypes of IP 3R were expressed in cultured SMC. DNA sequencing indicated that amplified products of IP 3R Ⅱ subtype was identical to reported sequence from gene bank. Conclusion Our results suggested that RT-PCR established by us can be used as a reliable method for investigating expression of mRNA for IP 3R subtypes.

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刘乃丰,董莉,何家声.大鼠组织中三磷酸肌醇受体亚型基因的表达[J].中国动脉硬化杂志,1999,7(3):201~204.

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  • 收稿日期:1999-04-27
  • 最后修改日期:1999-08-05
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