Ribulose-l,5-bisphosphate carboxylase small subunit gene (rbcS) is present with multi-gene family in plant genome. In Glycine max, the rbcS polypeptide (EC4.1.1.39) is encoded by a gene family containing 4-8 members. ...Ribulose-l,5-bisphosphate carboxylase small subunit gene (rbcS) is present with multi-gene family in plant genome. In Glycine max, the rbcS polypeptide (EC4.1.1.39) is encoded by a gene family containing 4-8 members. Three full-length rbcS cDNA clones were isolated and characterized from soybean seedlings, and both of their nucleotide and amino acid sequences showed high similarity. Differential accumulation of the rbcS mRNA was observed among roots, hypocotyls, cotyledons, epicotyls and leaves. The rbcS genes were up-regulated by various external factors such as salicylic acid (SA), salt stress and drought stress. The expression level of rbcS genes after being treated by 2.0 mmol/L SA and0.4% NaCl, respectively, is 2.5-3.0-fold as high as that of control sample. Moreover, soybean rbcS mRNA was accu-mulated with diurnal variation but easily influenced by light and low temperature.展开更多
A structural gene with 385 bp, which codes for the mature small subunit (rbcS) of ribulose-1,5-bi9phosphate carboxylase/oxygenase from tobacco, has been redesigned according to the codon usage of E.coli and chemically...A structural gene with 385 bp, which codes for the mature small subunit (rbcS) of ribulose-1,5-bi9phosphate carboxylase/oxygenase from tobacco, has been redesigned according to the codon usage of E.coli and chemically synthesized. To facilitate the systematic investigation of structure-functional relationship of the rbcS by site-specific mutagenesis, twenty one unique restriction sites distributed throughout the synthetic gene were designed. Gene synthesis was started from chemically synthesizing sixteen oligonucleotides each with 23-66 nucleotides, and these oligonu-cleotides were annealed to form eight duplexes from which 5'- and 3'- two half molecules were formed by stepwise T4 DNA ligase reaction, and then each half molecule was cloned into plasmid pWR13, after that, two half molecules were further confirmed by DNA sequencing, finally both half molecules excised from the cloning plasmid were recombinated to form plasmid pCOTrbcS containing the whole structural gene for tobacco rbcS.展开更多
[Objective] The aim was to study adaptive evolution of the large subunits of RubisCO in Magnoliophyta crops. [Method] Taking Magnoliophyta crops such as corn and rice as research materials, the analysis on molecular a...[Objective] The aim was to study adaptive evolution of the large subunits of RubisCO in Magnoliophyta crops. [Method] Taking Magnoliophyta crops such as corn and rice as research materials, the analysis on molecular adaptive evolution was carded out by using codon replacement and maximum likelihood methods. [ Result] The RubisCO suffered positive selection effect and six amino acid sites were identified. [ Conclusion] The six amino acid sites are of important guiding significance for studying catalytic activity of RubisCO large subunits and crop improvement.展开更多
基金This work was supported by the National Key Basic Research Special Funds of China (Grant No. G1998010209).
文摘Ribulose-l,5-bisphosphate carboxylase small subunit gene (rbcS) is present with multi-gene family in plant genome. In Glycine max, the rbcS polypeptide (EC4.1.1.39) is encoded by a gene family containing 4-8 members. Three full-length rbcS cDNA clones were isolated and characterized from soybean seedlings, and both of their nucleotide and amino acid sequences showed high similarity. Differential accumulation of the rbcS mRNA was observed among roots, hypocotyls, cotyledons, epicotyls and leaves. The rbcS genes were up-regulated by various external factors such as salicylic acid (SA), salt stress and drought stress. The expression level of rbcS genes after being treated by 2.0 mmol/L SA and0.4% NaCl, respectively, is 2.5-3.0-fold as high as that of control sample. Moreover, soybean rbcS mRNA was accu-mulated with diurnal variation but easily influenced by light and low temperature.
基金Project supported by grants from the High Technology Development Program of China,From the National Key Laboratory of Biooganic and Natural Product Chemistry of China,and from the Laboratory of Computer Chemistry,Chinese Academy of Sciences.
文摘A structural gene with 385 bp, which codes for the mature small subunit (rbcS) of ribulose-1,5-bi9phosphate carboxylase/oxygenase from tobacco, has been redesigned according to the codon usage of E.coli and chemically synthesized. To facilitate the systematic investigation of structure-functional relationship of the rbcS by site-specific mutagenesis, twenty one unique restriction sites distributed throughout the synthetic gene were designed. Gene synthesis was started from chemically synthesizing sixteen oligonucleotides each with 23-66 nucleotides, and these oligonu-cleotides were annealed to form eight duplexes from which 5'- and 3'- two half molecules were formed by stepwise T4 DNA ligase reaction, and then each half molecule was cloned into plasmid pWR13, after that, two half molecules were further confirmed by DNA sequencing, finally both half molecules excised from the cloning plasmid were recombinated to form plasmid pCOTrbcS containing the whole structural gene for tobacco rbcS.
文摘[Objective] The aim was to study adaptive evolution of the large subunits of RubisCO in Magnoliophyta crops. [Method] Taking Magnoliophyta crops such as corn and rice as research materials, the analysis on molecular adaptive evolution was carded out by using codon replacement and maximum likelihood methods. [ Result] The RubisCO suffered positive selection effect and six amino acid sites were identified. [ Conclusion] The six amino acid sites are of important guiding significance for studying catalytic activity of RubisCO large subunits and crop improvement.
