新型类固醇磺基转移酶的分析-猫眼课题宝

新型类固醇磺基转移酶的分析

批准号：

15590965

负责人：

SHIMIZU Chikara

金额：

$2.24万

依托单位：

HOKKAIDO UNIVERSITY

依托单位国家：

日本

项目类别：

Grant-in-Aid for Scientific Research (C)

财政年份：

2003

资助国家：

日本

项目状态：

已结题

起止时间：

2003 至 2004

来源：

https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590965/

关键词：

sulfotransferase transcription in situ hybridization immunohistochemistry promoter

项目摘要

项目成果

SHIMIZU Chikara的其他基金

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中文摘要

翻译

在这项研究中，我旨在阐明新发现的小鼠类固醇/甾醇硫转移酶（SULT2B1）的组织分布和转录调控。作为第一步，进行原位杂交分析。SULT2B1a是由SULT2B1基因编码的同工酶，主要在中枢神经系统中表达，而SULT2B1b mRNA在皮肤中检测到。特异性多克隆抗体的免疫组织化学分析证实了这一点。为了分析SULT2B1基因的转录调控，我分离了SULT2B1a和2B1b转录起始位点上游约2kb的基因组DNA片段，并将片段连接到荧光素酶表达载体上。由于在小鼠角质化鳞状细胞癌Pam212细胞系中检测到SULT2B1b mRNA，因此我将2 kb的启动子/增强子荧光素酶构建体转染到细胞中并分析了荧光素酶的活性。结果表明，含有SULT2B1b 5‘侧链区域的报告基因构建体荧光素酶活性比空载体和反向dna连接载体高10倍，表明该2-kb的5’侧链区域含有启动子和/或增强子结合序列。此外，我还利用PCR和退化引物克隆了兔SULT2B1b全长cDNA。RT-PCR实验显示，兔SULT2B1b mRNA在动脉中表达。生成兔SULT2B1b蛋白的多克隆抗体，在蛋白水平上确认表达。SULT2B1b与动脉粥样硬化发生之间的关系是我的下一个重点。

英文摘要

In this research, I aimed to clarify tissue distributions and transcriptional regulation of newly identified mouse steroid/sterol sulfotransferase (SULT2B1). As the first step, in situ hybridization analysis was performed. Message of SULT2B1a, an isozyme encoded by SULT2B1 gene, is expressed mainly in central nervous system, while SULT2B1b mRNA is detected in skin. This was confirmed by immmunohistochemical analysis using specific polyclonal antibodies. To analyze transcriptional regulation of SULT2B1 gene, I isolated approximately 2-kb genomic DNA fragments upstream of transcription start sites of SULT2B1a and 2B1b and ligated the fragments to luciferase expression vector. Because SULT2B1b mRNA is detected in Pam212 cell line derived from mouse keratinizing squamous cell carcinoma, I transfected 2-kb promoter/enhancer luciferase construct into the cells and analyzed the luciferase activity. The luciferase activity of reporter construct containing SULT2B1b 5' flanking region showed 10-fold higher than those of empty vector and reversely-oriented DNA-ligated vector, revealing that this 2-kb 5'-flanking region contains promoter and/or enhancer binding sequence. Furthermore, I have cloned rabbit SULT2B1b full-length cDNA using PCR and degenerated primers. RT-PCR experiment revealed that rabbit SULT2B1b mRNA is expressed in artery. Generating polyclonal antibody against rabbit SULT2B1b protein, I will confirm the expression at protein level. The association between SULT2B1b and atherosclerogenesis is my next focus.

期刊论文(70)

专著(0)

科研奖励(0)

会议论文

A rare case of Gitelman's syndrome presenting with hypocalcemia and osteopenia.

吉特尔曼综合征的罕见病例，表现为低钙血症和骨质减少。

DOI： --

发表时间：

期刊：

J Endocrinol Invest (in press)

影响因子： --

作者：

[Shimizu C et al., Shimizu C et al., Nagai S et al., Kubo M et al., Suzuki F et al., Ishizu A et al., Kijima H et al., 石井伸明他, Shimizu C et al., Nagai S et al., Kubo M et al., Suzuki F et al., Ishizu A et al., Kijima H et al., Ishizu A et al., Kijima H et al., Nagai S et al., Suzuki F et al., 石井伸明ほか, Shimizu C et al., Nakamura A et al., Nagai S et al., Matsumoto R et al., Lee YC et al., Nakamura A et al.]

通讯作者： Nakamura A et al.

A case of reversed pituitary dysfunction with intrasellar mass.

伴有鞍内肿块的逆转垂体功能障碍一例。

DOI： --

发表时间：

期刊：

J Endocrinol Invest (in press)

影响因子： --

作者：

通讯作者： Nagai S et al.

Adulut-onset Idiopathic Hypogonadotropic Hypogonadism due to Isolated Pituitary Gonadotropin Deficiency

孤立性垂体促性腺激素缺乏所致的成人特发性低促性腺激素性性腺功能减退症

DOI： --

发表时间： 2004

期刊：

Intern Med 43

影响因子： --

作者：

[Shimizu C et al., Shimizu C et al., Nagai S et al., Kubo M et al., Suzuki F et al.]

通讯作者： Suzuki F et al.

Suzuki F: "Adulut-onset Idiopathic Hypogonadotropic Hypogonadism due to Isolated Pituitary Gonadotropin Deficiency"Internal Medicine. (in press). (2004)

铃木 F：“由于孤立性垂体促性腺激素缺乏所致的成人特发性低促性腺激素低下症”内科。

DOI： --

发表时间：

期刊：

影响因子： --

作者：

[]

通讯作者：

Identification of a functional peroxysonie proliferator-activated receptor responsive element within the murine perilipin gene.

小鼠周脂质基因内功能性过氧化氢增殖物激活受体反应元件的鉴定。

DOI： --

发表时间： 2004

期刊：

Endocrinology 145

影响因子： --

作者：

[Shimizu C et al., Shimizu C et al., Nagai S et al., Kubo M et al., Suzuki F et al., Ishizu A et al., Kijima H et al., 石井伸明他, Shimizu C et al., Nagai S et al.]

通讯作者： Nagai S et al.