title: Molecular cloning, sequence analysis and expression in Escherichia coli of Camelus dromedarius glucose-6-phosphate dehydrogenase cDNA.
abstract: This study determined the full length sequence of glucose-6-phosphate dehydrogenase cDNA (G6PD) from the Arabian camel Camelus dromedarius using reverse transcription polymerase chain reaction. The C. dromedarius G6PD has an open reading frame of 1545 bp, and the cDNA encodes a protein of 515 amino acid residues with a molecular weight of 59.0 KDa. The amino acid sequence showed the highest identity with Equus caballus (92%) and Homo sapiens (92%). The G6PD cDNA was cloned and expressed into Escherichia coli as a fusion protein and was purified in a single chromatographic step using nickel affinity gel column. The purity and the molecular weight of the enzyme were checked on SDS-PAGE and the purified enzyme showed a single band on the gel with a molecular weight of 63.0 KDa. The specific activity of G6PD was determined to be 289.6 EU/mg protein with a fold purification of 95.45 and yield of 56.8%.
authors.affiliation: The Genome Research Chair, Biochemistry Department, College of Science, King Saud University, Bldg 5, Lab AA10, PO Box 2454, Riyadh, Saudi Arabia. hsaeed1@ksu.edu.sa
authors.forename: Hesham Mahmoud
authors.initials: HM
authors.lastname: Saeed
authors.forename: Mohammad Saud
authors.initials: MS
authors.lastname: Alanazi
authors.forename: Zainularifeen
authors.initials: Z
authors.lastname: Abduljaleel
authors.forename: Abdullah
authors.initials: A
authors.lastname: Al-Amri
authors.forename: Zahid
authors.initials: Z
authors.lastname: Khan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002162:Camelus / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D005954:Glucosephosphate Dehydrogenase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D015151:Immunoblotting; D008297:Male; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein
chemical_list: D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D005954:Glucosephosphate Dehydrogenase