Genetic and functional analysis of myosin I of blood cells

血细胞肌球蛋白 I 的遗传和功能分析

• 批准号: 09671090
• 负责人: HIGASHIHARA Masaaki
• 金额: $ 1.73万
• 依托单位: KITASATO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671090/
• 关键词: myosin; myosin I; cloning; platelet; cord blood; sequence; cDNA; library; mpl; 網血小板; 血液細胞; モノクローナル抗体

We had made the cDNA libraries of human cord blood cells, K562 (erythroleukemia cell line) and Meg-01 (megakaryocytic cell line). Furthermore, we established the method of calculation of "retiplatelet" for the evaluation of megakaryocyte-platelet production, using flow cytometric analysis. To make cloning of novel myosin, we made random primer of myosin I. Finally we had got the 4 clones using K562 library as template. By homology-analysis we found one is myosin VI, another myosin IX, and two unknown clones. RT-PCR method revealed that one of these two clones was also found in CMK and Meg-01, both of which are megakaryocytic cell lines. These results suggest that this clone is a novel unconventional myosin. This novel myosin is constituted in cultured cell from human organs including peripheral blood cells and bone marrow cells.We are now searching new myosin using the library of human cord blood cells, about 10,000 clones of which were already sequenced.Another our projects 1) sequencing both 20-kDa and 17-kDa myosin light chains, 2) functional analysis of conventional myosin heavy chain IIA and IIB in blood cells, are also underway.

我们构建了人脐血细胞、红白血病细胞系K562和巨核细胞系Meg-01的cDNA文库。此外，我们还建立了流式细胞术分析巨核细胞血小板生成的“血小板网”计算方法。为了克隆新的肌球蛋白，我们设计了肌球蛋白I的随机引物。最后以K562文库为模板得到了4个克隆。通过同源性分析，我们发现一个是肌球蛋白VI，另一个肌球蛋白IX，和两个未知的克隆。RT-PCR方法检测发现，CMK和Meg-01巨核细胞系中也存在这两个克隆。这些结果表明，该克隆是一种新的非常规肌球蛋白。这种新的肌球蛋白是在培养的人体器官细胞中形成的，包括外周血细胞和骨髓细胞。我们现在正在利用人脐带血细胞库寻找新的肌球蛋白，其中大约10，000个克隆已经测序。另一个我们的项目1）对20-kDa和17-kDa肌球蛋白轻链进行测序，2）常规肌球蛋白重链IIA和IIB在血细胞中的功能分析也在进行中。

项目成果

Horie R et al.: "Frequent expression of the variant CD30"Am J. Pathol. (in press).

Horie R 等人：“变体 CD30 的频繁表达”Am J. Pathol。

Koike Y., Yoneyama A., Shirai J, Ishida T., Shoda E., Miyazaki K., Sunaga S., Horie R., Aoki K., Koike K., Ogata I., Tahara T., Kato T., Nakahara K., Kariya T., Higashihara M.: "Evaluation of thrombopoiesis in thrombocytopenic disorders by simultaneous me

小池 Y.、米山 A.、白井 J、石田 T.、正田 E.、宫崎 K.、须永 S.、堀江 R.、青木 K.、小池 K.、绪方 I.、田原 T.、加藤 T.

Horie R, Yatomi Y, Wakabayashi T, Ohno Y, Eriguchi M, Higashihara M, Nakahara K, Watanabe T: "Primary gastric T-cell lymphomas : Report of two cases and a review of the literature"Jpn J Clin Oncol. 29. 171-178 (1999)

Horie R、Yatomi Y、Wakabayashi T、Ohno Y、Eriguchi M、Higashihara M、Nakahara K、Watanabe T：“原发性胃 T 细胞淋巴瘤：两例报告和文献综述”Jpn J Clin Oncol。

Koike Y et al.: "Evaluation of thrombopoiesis in thrombocytopenic"Thromb, Haemost. 79. 1106-1110 (1998)

Koike Y 等人：“血小板减少症中血小板生成的评估”Thromb，Haemost。

Koike Y et al: "Evaluation of thrombopoiesis in thrombocytopenic disorclers" Thromb.Haemostalsis. 79. 1106-1110 (1998)

Koike Y 等人：“血小板减少症紊乱者中血小板生成的评估”Thromb.Haemostalsis。