CHARACTERIZATION OF THE 5' REGULATORY REGION OF THE MOUSE (M) CGTP1 GENE

小鼠 (M) CGTP1 基因 5 调控区的特征

• 批准号: 5203496
• 负责人: M NAPOLITANO
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5203496
• 关键词: 3'5' cyclic nucleotide phosphodiesterase; 3T3 cells; Sf9 cell line; adipocytes; animal genetic material tag; complementary DNA; cyclic GMP; gene expression; genetic regulation; human genetic material tag; laboratory mouse; molecular cloning; northern blottings; open reading frames; phosphodiesterase inhibitors; recombinant DNA; regulatory gene

Seven cyclic nucleotide phosphodiesterase (PDE) gene families (Types I- VII) have been identified. We have cloned and expressed representatives of two sub-types of Type III cGMP-inhibited (cGI) PDEs, rat (R) and Human (H) cGIP1 and 2. A membrane-associated adipocyte cGI PDE (McGIP1 isoform) appears during differentiation of 3T3-L1 adipocytes. To learn more about the regulation of McGIP1 gene expression we cloned the cDNA for the cGI-PDE from a mouse adipocyte lambda gt 11 cDNA library (McGIP1 cDNA) The open reading frame of 3303 nucleotides is very similar to the sequence of RcGIP1 cDNA; the carboxyl-terminal region includes the putative catalytic domain highly conserved among mammalian PDEs. On Northern blots, McGIP1 cDNA hybridizes with a prominent approx. 5.5-kb band in poly(A)+ RNA from mouse adipose tissue, testis, liver, lung, and cerebellum, but not heart. Active recombinant McGIP1 was expressed in Sf9 cells transfected with recombinant baculovirus vector.

七个环核苷酸磷酸二酯酶 (PDE) 基因家族（I 型- VII) 已确定。 我们克隆并表达了代表 III 型 cGMP 抑制 (cGI) PDE 的两种亚型，大鼠 (R) 和 人 (H) cGIP1 和 2. 膜相关脂肪细胞 cGI PDE (McGIP1 isoform）出现在 3T3-L1 脂肪细胞的分化过程中。 为了了解更多有关 McGIP1 基因表达调控的信息，我们克隆了 来自小鼠脂肪细胞 lambda gt 11 cDNA 的 cGI-PDE cDNA 文库（McGIP1 cDNA）3303个核苷酸的开放阅读框是 与RcGIP1 cDNA的序列非常相似；羧基末端 区域包括高度保守的推定催化结构域 哺乳动物 PDE。 在 Northern 印迹中，McGIP1 cDNA 与 突出约。来自小鼠脂肪组织的 Poly(A)+ RNA 中的 5.5-kb 条带， 睾丸、肝脏、肺和小脑，但不是心脏。 活性重组 McGIP1在重组体转染的Sf9细胞中表达 杆状病毒载体。