Vora S, Hong F, Olender E
Biochem Biophys Res Commun. 1986 Mar 13;135(2):615-21. doi: 10.1016/0006-291x(86)90037-9.
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 approximately 2000 bp in length. It represents approximately 1350 bp of the C-terminus coding sequence of the enzyme, approximately 500 bp of the 3'-untranslated region and approximately 150 bp of the vector sequences. The identity of the pO4 cDNA was established by the observation of a high degree of homology (approximately 95%) between the deduced amino acid sequence with the published protein sequence of rabbit muscle 6-phosphofructokinase, and 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.
已从一个使用冈本-伯格方法构建的人成纤维细胞cDNA文库中分离出编码人肌肉6-磷酸果糖激酶（EC.2.7.1.11）的cDNA。作为pcD重组体pO4的Bam H1片段分离出的cDNA长度约为2000 bp。它代表该酶C端编码序列约1350 bp、3'-非翻译区约500 bp以及载体序列约150 bp。通过观察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（约95%），以及利用体细胞杂种将该序列定位到人染色体1（PFKM的已知位置），确定了pO4 cDNA的身份。基于免疫化学证据，我们之前不仅预测了脊椎动物肌肉磷酸果糖激酶在结构上有显著保守性，而且预测了所有三种脊椎动物磷酸果糖激酶同工酶之间存在部分结构同一性。因此，预计pO4 cDNA能够从多种脊椎动物物种中分离出肌肉和非肌肉磷酸果糖激酶的cDNA。
