Premium
cDNA cloning and expression of cysteine synthase B localized in chloroplasts of Spinacia oleracea
Author(s) -
Saito Kazuki,
Tatsuguchi Kazuyo,
Murakoshi Isamu,
Hirano Hisashi
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)80127-g
Subject(s) - spinacia , complementary dna , biology , microbiology and biotechnology , biochemistry , transit peptide , cdna library , open reading frame , cysteine , peptide sequence , expression vector , spinach , atp synthase , nucleic acid sequence , escherichia coli , chloroplast , gene , enzyme , recombinant dna , plastid
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.