Identification, chromosomal assignment, and expression analysis of the human homeodomain-containing gene Orthopedia (OTP).
Identification, chromosomal assignment, and expression analysis of the human homeodomain-containing gene Orthopedia (OTP).
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含有人类同源结构域的基因 Orthopedia (OTP) 的鉴定、染色体分配和表达分析。
DOI：
10.1006/geno.1999.5882
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发表时间：
1999
期刊：
影响因子：
4.4
通讯作者：
Leckman,JF
中科院分区：
文献类型：
--
作者：
Lin,X;State,MW;Vaccarino,FM;Greally,J;Hass,M;Leckman,JF
同源结构域 (HD) 基因是螺旋-转角-螺旋转录因子，在细胞命运的规范中发挥关键作用。在中枢神经系统 (CNS) 中，HD 基因不仅可以沿轴定位细胞，还可以指定细胞迁移模式，并可能影响轴突连接。为了鉴定参与人类中枢神经系统发育的新 HD 基因，我们克隆、表征并绘制了小鼠 HD 基因 Orthopedia (Otp) 的人类同源物，该基因的产物存在于小鼠下丘脑、杏仁核和脑干内的多个细胞群中。使用源自小鼠 Otp 序列的探针筛选人类 cDNA 和基因组文库，以找到人类同源物 OTP。推导的人类 cDNA 开放阅读框的氨基酸序列与小鼠 Otp 具有 99% 同源性，与海胆和果蝇相比，显示出高度的保守性。使用辐射杂交图谱和荧光原位杂交将 OTP 定位到人类染色体 5q13.3。从含有 OTP 的 YAC 克隆中鉴定出侧翼标记。通过蛋白质印迹分析，使用针对 OTP 蛋白 C 末端保守的 13 个氨基酸序列产生的新型多克隆抗体，在 17 周人类胎儿脑组织中发现了单一推定的 OTP 基因产物。通过免疫组织化学证实了发育中的人类下丘脑中的表达。
Homeodomain (HD) genes are helix-turn-helix transcription factors that play key roles in the specification of cell fates. In the central nervous system (CNS), HD genes not only position cells along an axis, but also specify cell migration patterns and may influence axonal connectivity. In an effort to identify novel HD genes involved in the development of the human CNS, we have cloned, characterized, and mapped the human homologue of the murine HD gene Orthopedia (Otp), whose product is found in multiple cell groups within the mouse hypothalamus, amygdala, and brain stem. Human cDNA and genomic libraries were screened with probes derived from mouse Otp sequences to find the human homologue, OTP. The deduced amino acid sequence of the open reading frame of the human cDNA is 99% homologous to mouse Otp and demonstrates a high degree of conservation when compared to sea urchin and Drosophila. OTP was mapped to human chromosome 5q13.3 using radiation hybrid panel mapping and fluorescence in situ hybridization. Flanking markers were identified from YAC clones containing OTP. A single putative OTP gene product was found in 17-week human fetal brain tissue by Western blot analysis using a novel polyclonal antibody raised against a conserved 13-amino-acid sequence at the C-terminus of the OTP protein. Expression in the developing human hypothalamus was confirmed by immunohistochemistry.