cDNA cloning and expression of cysteine synthase B localized in chloroplasts of Spinacia oleracea.
The cDNA clones for cysteine synthase B, which is localized in chloroplasts of Spinacia oleracea L., were isolated by screening a library with synthetic oligonucleotides encoding a partial peptide sequence of the purified protein. Nucleotide sequence analysis revealed an open reading frame encoding a polypeptide of 383 amino acids containing a putative transit peptide of 52 amino acids. A bacterial expression vector of the cDNA clone could genetically complement an Escherichia coli auxotroph lacking cysteine synthase and could produce the functionally active and immuno-reactive cysteine synthase in E. coli. RNA blot hybridization suggested that the transcripts were primarily accumulated in leaves of spinach.
Saito
K
K
Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Tatsuguchi
K
K
Murakoshi
I
I
Hirano
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D002736:Chloroplasts / Q000201:enzymology*; D003001:Cloning, Molecular; D016384:Consensus Sequence; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant; D005816:Genetic Complementation Test; D008190:Lyases / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000737:chemistry; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014675:Vegetables / Q000235:genetics
D018076:DNA, Complementary; D015345:Oligonucleotide Probes; D010940:Plant Proteins; D008190:Lyases; C022706:S-sulfocysteine synthase