PCR CLONING OF NOVEL TYROSINE PHOSPHATASES FROM RHESUS MONKEY TISSUES

酪氨酸磷酸化酪氨酸激酶 信号蛋白中的残基参与了 胰岛素与胰岛素受体结合后的作用。 人民和 患有2型糖尿病的动物对胰岛素的作用有抵抗力。 治疗2型糖尿病的一种方法是提高胰岛素水平， 通过抑制参与的酶， 去磷酸化酪氨酸（酪氨酸磷酸酶）， 信号φ 为了鉴定潜在的酪氨酸磷酸酶 靶点药物，将抑制这些酶的活性， 需要鉴定在灵长类动物中起作用的特异性磷酸酶。 灵长类酪氨酸磷酸酶以前没有被克隆。 目的获得胰岛素敏感组织（脂肪、肌肉和 肝）用于酪氨酸的聚合酶链反应（PCR）克隆 磷酸酶 克隆由Sugen，Inc.结果 在加州地区灵长类动物研究中心收集组织 中心（CRPRC）。 从5名正常体重和5名 肥胖的成年雄性恒河猴。 脂肪（0.5 g）和肌肉（0.5 g）样品， 在氯胺酮麻醉下通过手术活检收集。 肝脏样品 （20-50 mg）通过超声引导下穿刺活检收集， 氯胺酮麻醉 由CRPRC进行活检程序 兽医 将样品置于无菌塑料小瓶中， 立即在液氮中冷冻。 冷冻组织样本 用干冰运输至Sugen，Inc.在加利福尼亚州的红杉城。 苏根有 从样品中克隆了几种酪氨酸磷酸酶。 组织样本 在本项目/基因阵列方案下也获得了 我们实验室的分析。 未来的方向没有进一步的实验 然而，在未来， 我们需要从这些或其他动物身上获取更多的组织 类似的实验。 关键词酪氨酸磷酸酶，肥胖， 糖尿病，PCR克隆，肌肉，脂肪，肝脏基金Sugen，Inc.， 红杉城，加利福尼亚州

Significance Tyrosine kinases which phosphorylate tyrosine residues in signaling proteins are involved in transduction of insulin's actions after it binds to insulin receptors. People and animals with Type 2 diabetes are resistant to the actions of insulin. One approach to treating Type 2 diabetes is to enhance the insulin signal by inhibiting the enzymes which are involved in dephosphorylating tyrosine (Tyrosine phosphatases) which inactivates the signal. In order to identify potential tyrosine phosphatase targets for drugs that will inhibit the activity of these enzymes the specific phosphatases functioning in primates need to be identified. Primate tyrosine phosphatases have not previously been cloned. Objectives To obtain insulin sensitive tissues (fat, muscle, and liver) for polymerase chain reaction (PCR) cloning of tyrosine phosphatases. Cloning was performed by and at Sugen, Inc. Results Tissues were collected at the California Regional Primate Research Center (CRPRC). Samples were obtained from 5 normal weight and 5 obese adult male rhesus. Fat (0.5 g) and muscle (0.5 g) samples was collected by surgical biopsy under ketamine anesthesia. Liver samples (20-50 mg) were collected by ultrasound-guided needle biopsy under ketamine anesthesia. Biopsy procedures were performed by CRPRC veterinarians. Samples were placed in sterile plastic vials and immediately frozen in liquid nitrogen. The frozen tissue samples were shipped on dry ice to Sugen, Inc. in Redwood City, CA. Sugen has cloned several tyrosine phosphatases from the samples. Tissue samples have also been obtained under this project/protocol for gene array analysis by our laboratory. Future Directions No further experiments are planned at this time, however, it is possible that in the future we will want to obtain additional tissues from these or other animals for similar experiments. KEY WORDS tyrosine phosphatases, obesity, diabetes, PCR cloning, muscle, Adipose, liver FUNDING Sugen, Inc., Redwood City, CA