Isolation and characterization of the gene coding for cytosolic phosphoenolpyruvate carboxykinase (GTP) from the rat.
Isolation and characterization of the gene coding for cytosolic phosphoenolpyruvate carboxykinase (GTP) from the rat.
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大鼠胞质磷酸烯醇丙酮酸羧激酶 (GTP) 编码基因的分离和表征。
DOI：
10.1073/pnas.80.12.3656
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发表时间：
1983
影响因子：
11.1
通讯作者：
Hanson,RW
中科院分区：
文献类型：
--
作者：
Yoo-Warren,H;Monahan,JE;Short,J;Short,H;Bruzel,A;Wynshaw-Boris,A;Meisner,HM;Samols,D;Hanson,RW
从噬菌体lambda Charon 4A中含有大鼠基因组的重组文库中分离到大鼠胞质磷酸烯醇丙酮酸羧激酶（GTP）基因[GTP：草酰乙酸羧解酶（转磷酸化），EC 4.1.32]。该分离克隆lambda PCK1包含磷酸烯醇丙酮酸羧激酶的完整基因，在5‘端和3’端分别约等于7千碱基和1千碱基。限制性内切酶定位、r环定位和部分DNA序列分析表明，该基因长度约为6.0 kb（编码2.8 kb的mRNA），含有8个内含子。用各种限制性内切酶消化的大鼠DNA的Southern印迹显示了从lambda PCK1限制性内切图预测的模式。分离出该基因5'端包含1.2 kb限制性片段的控制区，并将其亚克隆到pBR322中。该基因的这一部分包含通常的转录起始序列和一个24碱基序列，几乎与人类原皮皮激素基因5'侧区发现的序列相同，已知该基因受糖皮质激素调节。磷酸烯醇丙酮酸羧激酶基因的1.2 kb片段可以通过体外转录试验转录成预测大小的独特RNA片段。
The gene for cytosolic phosphoenolpyruvate carboxykinase (GTP) [GTP:oxaloacetate carboxy-lyase (transphosphorylating), EC 4.1.1.32] from the rat was isolated from a recombinant library containing the rat genome in phage lambda Charon 4A. The isolated clone, lambda PCK1, contains the complete gene for phosphoenolpyruvate carboxykinase and approximately equal to 7 kilobases (kb) of flanking sequence at the 5' end and 1 kb at the 3' terminus. Restriction endonuclease mapping, R-loop mapping, and partial DNA sequence assay indicate that the gene is approximately equal to 6.0 kb in length (coding for a mRNA of 2.8 kb) and contains eight introns. Southern blotting of rat DNA digested with various restriction enzymes shows a pattern predicted from the restriction map of lambda PCK1. A control region at the 5' end of the gene contained in a 1.2-kb restriction fragment was isolated and subcloned into pBR322. This segment of the gene contains the usual transcription start sequences and a 24-base sequence virtually identical to the sequence found in the 5'-flanking region of the human proopiomelonocortin gene, which is known to be regulated by glucocorticoids. The 1.2-kb fragment of the phosphoenolpyruvate carboxykinase gene can be transcribed into a unique RNA fragment of predicted size by an in vitro transcription assay.