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
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
Ulva prolifera is a typical green alga in subtidal areas and can grow tremendously fast. A highly efficient Rubisco enzyme which is encoded by Up Rbc L gene may contribute to the rapid growth. In this study, the full-...Ulva prolifera is a typical green alga in subtidal areas and can grow tremendously fast. A highly efficient Rubisco enzyme which is encoded by Up Rbc L gene may contribute to the rapid growth. In this study, the full-length Up Rbc L open reading frame(ORF) was identified, which encoded a protein of 474 amino acids. Phylogenetic analysis of Up Rbc L sequences revealed that Chlorophyta had a closer genetic relationship with higher plants than with Rhodophyta and Phaeophyta. The two distinct residues(aa11 and aa91) were presumed to be unique for Rubisco catalytic activity. The predicted three-dimensional structure showed that one α/β-barrel existed in the C-terminal region, and the sites for Mg^(2+)coordination and CO_2 fixation were also located in this region. Gene expression profile indicated that Up Rbc L was expressed at a higher level under light exposure than in darkness. When the culture temperature reached 35℃, the expression level of Up Rbc L was 2.5-fold lower than at 15℃, and the carboxylase activity exhibited 13.8-fold decrease. Up Rbc L was heterologously expressed in E. coli and was purified by Ni^(2+) affinity chromatography. The physiological and biochemical characterization of recombinant Rubisco will be explored in the future.展开更多
Species of the red algal order Acrochaetiales mostly inhabit marine environments,with only two freshwater taxa Audouinella and Ottia.A new genus and species are described for freshwater red alga Pauciramus yunnanensis...Species of the red algal order Acrochaetiales mostly inhabit marine environments,with only two freshwater taxa Audouinella and Ottia.A new genus and species are described for freshwater red alga Pauciramus yunnanensis from Ailao Mountain,Yunnan,China.It is closely related to Ottia and a new order Ottiales was proposed for these genera.Pauciramus has unique combination of morphological characteristics including the following:plants caespitose and densely pulvinate,slender uniseriate filaments with well-developed rhizoids,rarely branched,cylindrical vegetative cell with a single,ribbon-shaped and parietal chloroplast,reproduction by tetrasporangia,and dense sporangial branchlet only at the upper portion of filaments.Phylogenetic analysis of sequence data from the plastid ribulose-1,5-bisphosphate carboxylase-oxygenase large-subunit(rbc L),small subunit gene of the ribosomal cistron(SSU)and barcode region near the 5′end of the mitochondrial cytochrome oxidase subunit I(COI-5P)indicated that:the new taxon,P.yunnanensis,was in a well-supported clade with Ottia meiospora,and this clade was sister to order Palmariales and Acrochaetiales.To adhere to the principle of monophyly,a new freshwater order Ottiales including Ottia and Pauciramus is proposed.Despite the high sequence interspecific divergences and obvious morphological differences between genera Ottia and Pauciramus,seems impractical to establish a new family for a monospecific genus.Therefore,we temporarily classified Pauciramus into the family Ottiaceae,and made necessary revisions to the description to accommodate this genus.展开更多
Although a considerable increase in our knowledge concerning the importance of metabolic adjustments to unfavorable growth conditions has been recently provided, relatively little is known about the adjustments which ...Although a considerable increase in our knowledge concerning the importance of metabolic adjustments to unfavorable growth conditions has been recently provided, relatively little is known about the adjustments which occur in response to fluctuation in environmental factors. Evaluating the metabolic adjustments occurring under changing environmental conditions thus offers a good opportunity to increase our current understanding of the crosstalk between the major pathways which are affected by such conditions. To this end, plants growing under normal conditions were transferred to different light and temperature conditions which were anticipated to affect (amongst other processes) the rates of photosynthesis and photorespiration and characterized at the physiological, molecular, and metabolic levels following this transition. Our results revealed similar behavior in response to both treatments and imply a tight connec- tivity of photorespiration with the major pathways of plant metabolism. They further highlight that the majority of the regulation of these pathways is not mediated at the level of transcription but that leaf metabolism is rather pre-poised to adapt to changes in these input parameters.展开更多
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.
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financially supported by the Scientific and Technical Supporting Programs of China(2008 BAC49B01)State Ocean Administration Project(200805069)
文摘Ulva prolifera is a typical green alga in subtidal areas and can grow tremendously fast. A highly efficient Rubisco enzyme which is encoded by Up Rbc L gene may contribute to the rapid growth. In this study, the full-length Up Rbc L open reading frame(ORF) was identified, which encoded a protein of 474 amino acids. Phylogenetic analysis of Up Rbc L sequences revealed that Chlorophyta had a closer genetic relationship with higher plants than with Rhodophyta and Phaeophyta. The two distinct residues(aa11 and aa91) were presumed to be unique for Rubisco catalytic activity. The predicted three-dimensional structure showed that one α/β-barrel existed in the C-terminal region, and the sites for Mg^(2+)coordination and CO_2 fixation were also located in this region. Gene expression profile indicated that Up Rbc L was expressed at a higher level under light exposure than in darkness. When the culture temperature reached 35℃, the expression level of Up Rbc L was 2.5-fold lower than at 15℃, and the carboxylase activity exhibited 13.8-fold decrease. Up Rbc L was heterologously expressed in E. coli and was purified by Ni^(2+) affinity chromatography. The physiological and biochemical characterization of recombinant Rubisco will be explored in the future.
基金Supported by the National Natural Science Foundation of China (No.41871037 to Shulian XIE and No.31800172 to Fangru NAN)the Fund for Shanxi"1331 Project"
文摘Species of the red algal order Acrochaetiales mostly inhabit marine environments,with only two freshwater taxa Audouinella and Ottia.A new genus and species are described for freshwater red alga Pauciramus yunnanensis from Ailao Mountain,Yunnan,China.It is closely related to Ottia and a new order Ottiales was proposed for these genera.Pauciramus has unique combination of morphological characteristics including the following:plants caespitose and densely pulvinate,slender uniseriate filaments with well-developed rhizoids,rarely branched,cylindrical vegetative cell with a single,ribbon-shaped and parietal chloroplast,reproduction by tetrasporangia,and dense sporangial branchlet only at the upper portion of filaments.Phylogenetic analysis of sequence data from the plastid ribulose-1,5-bisphosphate carboxylase-oxygenase large-subunit(rbc L),small subunit gene of the ribosomal cistron(SSU)and barcode region near the 5′end of the mitochondrial cytochrome oxidase subunit I(COI-5P)indicated that:the new taxon,P.yunnanensis,was in a well-supported clade with Ottia meiospora,and this clade was sister to order Palmariales and Acrochaetiales.To adhere to the principle of monophyly,a new freshwater order Ottiales including Ottia and Pauciramus is proposed.Despite the high sequence interspecific divergences and obvious morphological differences between genera Ottia and Pauciramus,seems impractical to establish a new family for a monospecific genus.Therefore,we temporarily classified Pauciramus into the family Ottiaceae,and made necessary revisions to the description to accommodate this genus.
文摘Although a considerable increase in our knowledge concerning the importance of metabolic adjustments to unfavorable growth conditions has been recently provided, relatively little is known about the adjustments which occur in response to fluctuation in environmental factors. Evaluating the metabolic adjustments occurring under changing environmental conditions thus offers a good opportunity to increase our current understanding of the crosstalk between the major pathways which are affected by such conditions. To this end, plants growing under normal conditions were transferred to different light and temperature conditions which were anticipated to affect (amongst other processes) the rates of photosynthesis and photorespiration and characterized at the physiological, molecular, and metabolic levels following this transition. Our results revealed similar behavior in response to both treatments and imply a tight connec- tivity of photorespiration with the major pathways of plant metabolism. They further highlight that the majority of the regulation of these pathways is not mediated at the level of transcription but that leaf metabolism is rather pre-poised to adapt to changes in these input parameters.