Uchida K
Nagahama Institute for Biochemical Science, Oriental Yeast Company, Ltd., Shiga, Japan.
Arch Biochem Biophys. 1991 Aug 1;288(2):558-61. doi: 10.1016/0003-9861(91)90235-b.
A cDNA encoding the nonmuscle-specific (type B) subunit of phosphoglycerate mutase (PGAM-B) was isolated and characterized. A cDNA probe, synthesized by the polymerase chain reaction (PCR) from rat liver cell mRNA using mixed primers specific to the amino acid sequence of human PGAM-B, was used to screen a rat liver cell cDNA library. The identity of the cDNA was confirmed by amino acid sequence data for 24 peptides obtained by digesting the purified protein with three different endopeptidases. The coding region encoded a polypeptide composed of 253 amino acid (plus the initiator Met). RNA blot analysis showed a single mRNA species of 1.7 kilobases in rat liver cell. The deduced amino acid sequence of rat PGAM-B was identical to that of human PGAM-B except for only one substitution at position 251 near the carboxyl terminus (valine for the rat and alanine for the human).
分离并鉴定了编码磷酸甘油酸变位酶(PGAM-B)非肌肉特异性(B型)亚基的cDNA。使用针对人PGAM-B氨基酸序列的混合引物,通过聚合酶链反应(PCR)从大鼠肝细胞mRNA合成的cDNA探针,用于筛选大鼠肝细胞cDNA文库。通过用三种不同的内肽酶消化纯化蛋白获得的24个肽段的氨基酸序列数据,证实了该cDNA的身份。编码区编码一个由253个氨基酸组成的多肽(加上起始甲硫氨酸)。RNA印迹分析显示大鼠肝细胞中有一个1.7千碱基的单一mRNA种类。大鼠PGAM-B推导的氨基酸序列与人PGAM-B的序列相同,只是在羧基末端附近的第251位有一个替换(大鼠为缬氨酸,人为丙氨酸)。