Smock S L, Pan L C, Castleberry T A, Lu B, Mather R J, Owen T A
Department of Cardiovascular and Metabolic Diseases, Central Research Division, Pfizer, Inc., Eastern Point Road, Groton, CT, USA.
Gene. 1999 Sep 17;237(2):393-402. doi: 10.1016/s0378-1119(99)00323-6.
Northern blot analysis of human placental RNA using a probe to the 5' end of the human prostaglandin E(2) (PGE(2)) EP2 receptor subtype coding region revealed the existence of a high abundance, low molecular weight transcript. To investigate the origin of this transcript, and its possible relationship to the human EP2 mRNA, we have cloned and characterized the gene encoding the human PGE(2) EP2 receptor subtype, identified transcriptional initiation and termination sites in two tissues (spleen and thymus), and determined its chromosomal localization. The human EP2 gene consists of two exons separated by a large intron, utilizes a common initiation site in both spleen and thymus at 1113 bp upstream of the translation initiation site, and has 3' transcript termini at 1140 bp and 1149 bp downstream of the translation stop site in spleen and thymus respectively. Southern and fluorescence in situ hybridization analysis demonstrated the human EP2 gene to be a single copy gene located in band 22 of the long arm of chromosome 14 (14q22). Though our initial interest in this gene was to investigate potential differential splicing of the human EP2 gene in placenta, this work demonstrates that the atypical transcript observed in placenta probably arises from a distinct, yet related, gene. Knowledge of the sequence, structure, and transcription events associated with the human EP2 gene will enable a broader understanding of its regulation and potential role in normal physiology and disease.
利用针对人前列腺素E(2)(PGE(2))EP2受体亚型编码区5'端的探针,对人胎盘RNA进行Northern印迹分析,结果显示存在一种高丰度、低分子量的转录本。为了研究该转录本的起源及其与人EP2 mRNA的可能关系,我们克隆并鉴定了编码人PGE(2) EP2受体亚型的基因,确定了两个组织(脾脏和胸腺)中的转录起始和终止位点,并确定了其染色体定位。人EP2基因由两个外显子组成,中间隔着一个大的内含子,在脾脏和胸腺中均利用翻译起始位点上游1113 bp处的一个共同起始位点,在脾脏和胸腺中分别在翻译终止位点下游1140 bp和1149 bp处有3'转录末端。Southern印迹和荧光原位杂交分析表明,人EP2基因是位于14号染色体长臂22带(14q22)的单拷贝基因。尽管我们最初对该基因的兴趣在于研究人EP2基因在胎盘中潜在的差异剪接,但这项工作表明,在胎盘中观察到的非典型转录本可能来自一个不同但相关的基因。了解与人EP2基因相关的序列、结构和转录事件,将有助于更广泛地理解其在正常生理和疾病中的调控及潜在作用。