Isolation and characterization of an avian A1 adenosine receptor gene and a related cDNA clone.
Isolation and characterization of an avian A1 adenosine receptor gene and a related cDNA clone.
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禽类 A1 腺苷受体基因和相关 cDNA 克隆的分离和表征。
DOI：
10.1042/bj3070729
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Green,RD
中科院分区：
文献类型：
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作者：
Aguilar,JS;Tan,F;Durand,I;Green,RD
我们分离出了鸡 A1 腺苷受体的基因以及编码相同腺苷受体的 cDNA。该 cDNA 克隆从脂肪组织和心脏 cDNA 文库中分离出来，编码 324 个氨基酸的蛋白质，与哺乳动物 A1 腺苷受体具有 80% 的同一性。该 cDNA 在人胚肾 (HEK) 293 细胞中的瞬时表达表明，它编码一种结合 [3H]CCPA（2-氯-N6-[环戊基-2,3,4,5-3H]环戊基腺苷，一种特异性激动剂放射性配体，KD 为 5.6 +/- 2.4 nM）的蛋白质。与鸡 cDNA 和促黄体激素/绒毛膜促性腺激素受体的 cDNA 表明 A1 腺苷受体激动剂拮抗牛促黄体激素的环 AMP 升高作用。第一个克隆包含 5'-非翻译序列，其中包含不包含 TATA 框的推定启动子区域、内含子和 A1 腺苷受体 cDNA 的前三分之一编码序列。该部分基因组克隆的第二个内含子终止于第二个内含子。第二个部分基因组克隆包含编码序列的其余部分和单个外显子中的 3'-非翻译元件，因此，鸡 A1 腺苷受体基因在 5'-非翻译区中包含一个内含子，并且在编码序列中包含至少一个内含子。
We have isolated the gene for a chick A1 adenosine receptor along with a cDNA that codes for the same adenosine receptor. The cDNA clone was isolated from both adipose tissue and heart cDNA libraries and encodes a 324-amino acid protein with 80% identity with mammalian A1 adenosine receptors. Transient expression of the cDNA in human embryonic kidney (HEK) 293 cells shows that it encodes a protein that binds [3H]CCPA (2-chloro-N6-[cyclopentyl-2,3,4,5-3H]cyclopentyladenosine, a specific agonist radioligand, with a KD of 5.6 +/- 2.4 nM. Cyclic AMP measurements in HEK 293 cells co-transfected with the chick cDNA and a cDNA for a luteinizing hormone/choriogonadotropin receptor shows that A1 adenosine receptor agonists antagonize the cyclic AMP-elevating effect of bovine luteinizing hormone. Two partial genomic clones were isolated. The first contains 5′-untranslated sequence including a putative promoter region which does not contain a TATA box, an intron and the first third of the coding sequence of the A1 adenosine receptor cDNA. The coding sequence of this partial genomic clone terminates at a second intron. The second partial genomic clone contains the rest of the coding sequence and the 3′-untranslated elements in a single exon. Thus the chick A1 adenosine receptor gene contains one intron in the 5′-untranslated region and a minimum of one intron in the coding sequence.