Pegues J C, Chen L S, Gordon A W, Ding L, Coleman W G
Section on Pharmacology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.
J Bacteriol. 1990 Aug;172(8):4652-60. doi: 10.1128/jb.172.8.4652-4660.1990.
The rfaD gene encodes ADP-L-glycero-D-mannoheptose-6-epimerase, an enzyme required for the biosynthesis of the lipopolysaccharide precursor ADP-L-glycerol-D-mannoheptose. The precise localization of the rfaD gene on a 1.3-kilobase SspI-HpaI fragment is reported. The rfaD gene and the flanking regions were completely sequenced. The location of the rfaD gene on the physical map of the Escherichia coli chromosome was determined. Primer extension studies were used to define the regulatory region of the rfaD gene. The cloned rfaD gene directed the synthesis of a 37,000-dalton polypeptide in several in vivo and in vitro expression systems. N-terminal analysis of purified ADP-L-glycero-D-mannoheptose-6-epimerase confirmed the first 34-amino-acid sequence deduced from the nucleotide sequence of the rfaD gene coding region. The primary structure of the rfaD protein contains the sequence fingerprint for the ADP-binding beta alpha beta fold at the N terminus.
rfaD基因编码ADP-L-甘油-D-甘露庚糖-6-表异构酶，这是一种参与脂多糖前体ADP-L-甘油-D-甘露庚糖生物合成的酶。本文报道了rfaD基因在一个1.3千碱基的SspI-HpaI片段上的精确定位。对rfaD基因及其侧翼区域进行了全序列测定。确定了rfaD基因在大肠杆菌染色体物理图谱上的位置。采用引物延伸研究来确定rfaD基因的调控区域。在多个体内和体外表达系统中，克隆的rfaD基因指导合成了一种37,000道尔顿的多肽。对纯化的ADP-L-甘油-D-甘露庚糖-6-表异构酶进行的N端分析证实了从rfaD基因编码区核苷酸序列推导的前34个氨基酸序列。rfaD蛋白的一级结构在N端包含ADP结合β-α-β折叠的序列指纹。
