cDNA cloning and expression of cysteine synthase B localized in chloroplasts of Spinacia oleracea

FEBS Lett. 1993 Jun 21;324(3):247-52. doi: 10.1016/0014-5793(93)80127-g.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Chloroplasts / enzymology*
  • Cloning, Molecular
  • Consensus Sequence
  • DNA, Complementary / genetics
  • Gene Expression
  • Genes, Plant
  • Genetic Complementation Test
  • Lyases / genetics*
  • Molecular Sequence Data
  • Oligonucleotide Probes / chemistry
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Vegetables / genetics

Substances

  • DNA, Complementary
  • Oligonucleotide Probes
  • Plant Proteins
  • Lyases
  • S-sulfocysteine synthase

Associated data

  • GENBANK/D14722