Isolation of a cDNA for human muscle 6-phosphofructokinase.
Isolation of a cDNA for human muscle 6-phosphofructokinase.
复制标题
人肌肉 6-磷酸果糖激酶 cDNA 的分离。
DOI：
10.1016/0006-291x(86)90037-9
复制
发表时间：
1986
影响因子：
3.1
通讯作者：
Olender,E
中科院分区：
文献类型：
--
作者：
Vora,S;Hong,F;Olender,E
人肌肉6-磷酸果糖激酶（EC. 2.7. 1.11）已从使用Okayama-Berg方法制备的人成纤维细胞cDNA文库中分离。作为pcD重组体pO 4的BamH 1片段分离的cDNA长度为约2000 bp。它代表了该酶C端编码序列的约1350 bp，3′非翻译区的约500 bp和载体序列的约150 bp。pO 4 cDNA的同一性通过以下方法确定：a）观察到推导的氨基酸序列与已发表的兔肌肉6-磷酸果糖激酶蛋白质序列之间的高度同源性（> 95%），和B）使用体细胞杂交体将该序列分配到人1号染色体（PFKM的已知位置）。基于免疫化学证据，我们先前预测不仅脊椎动物肌肉PFK的显着结构保守性，但所有三种脊椎动物PFK同工酶之间的部分结构一致性。因此，预期pO 4 cDNA允许从多种脊椎动物物种中分离肌肉和非肌肉PFK的cDNA。
A cDNA for human muscle 6-phosphofructokinase (EC. 2.7. 1.11) has been isolated from a human fibroblast cDNA library made using the Okayama-Berg procedure. The cDNA isolated as a Bam H1 fragment of the pcD recombinant, pO4, is∼ 2000 bp in length. It represents∼ 1350 bp of the C-terminus coding sequence of the enzyme,∼ 500 bp of the 3′-untranslated region and∼ 150 bp of the vector sequences. The identity of the pO4 cDNA was established by a) the observation of a high degree of homology (∼ 95%) between the deduced amino acid sequence with the published protein sequence of rabbit muscle 6-phosphofructokinase, and b) the assignment of the sequence to human chromosome 1 (the known location of PFKM) by using somatic cell hybrids. Based on immunochemical evidence, we had previously predicted not only a remarkable structural conservation of the vertebrate muscle PFK, but also partial structural identity among all three vertebrate PFK isozymes. The pO4 cDNA is, therefore, expected to permit isolation of cDNAs for muscle and non-muscle PFKs from a wide variety of vertebrate species.