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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。
To observe the gene expression of inositol 1,4,5-trisphosphate receptor subtypes in rat myocardium, cerebellum and cultured aortic vascular smooth muscle cells, the total R N A of tissue was extracted by one-step method using guanidine isothiocyanate- RT-PCR and β-actin were used as the internal standard to detect the inositol triphosphate receptor subtypes m R N A expression. The Type I PICCR product was recombined, cloned and sequenced. Three subtypes of inositol 1,4,5-trisphosphate receptors were found in myocardium, cerebellum and smooth muscle cells, and their expression levels were higher in cerebellum. The amplified fragment of P C R was 525 bp (cerebellum) or 405 bp (myocardium and smooth muscle cell) of type Ⅰ inositol triphosphate receptor, 405 bp of inositol 1,4,5-triphosphate receptor, and the type Ⅲ inositol triphosphate receptor 169 bp. Sequence analysis revealed that the cloned inositol dinucleotide triphosphate cD N A sequence was consistent with the designed c D N A target, suggesting that the established reverse transcription-polymerase chain reaction method can reliably study inositol triphosphate Recipients of different subtypes of gene expression.