The biosynthesis and signaling of plant hormones play a critical role in almost all biological processes.It is well-documented that phytohormones cross-talk with each other.Epigenetic mechanisms were suggested to regu...The biosynthesis and signaling of plant hormones play a critical role in almost all biological processes.It is well-documented that phytohormones cross-talk with each other.Epigenetic mechanisms were suggested to regulate expression of downstream targets in hormone signaling pathways that help implement hormone functions.This new layer of complexities that integrate epigenetic information such as DNA methylation,chromatin remodeling,histone modification,microRNAs and siRNAs with plant hormone signaling and regulations of gene expression,has been gradually revealed.In this short review,the author tries to assemble recent progress to establish a molecular linkage between these two large and momentum research fields and also to help readers digest the literature.展开更多
The coding regions of Ag85B MPT-64, and ESAT-6 secreted proteins were cloned initially into the eu-karyotic expression vector pJW4303, then transformed to E. coli Top 10 strain for plasmid DNA extraction and further a...The coding regions of Ag85B MPT-64, and ESAT-6 secreted proteins were cloned initially into the eu-karyotic expression vector pJW4303, then transformed to E. coli Top 10 strain for plasmid DNA extraction and further analysis. Plasmids containing the right insertion were se-quenced to confirm their identity. COS7 cells were trans-fected with a mixture containing serially diluted plasmid DNA encoding three secreted proteins and Lipofectin (Gibco). The supernatants and pellets prepared from various cell lines were run on SDS-PAGE gel and the expression of these proteins in COS7 cells were demonstrated by immunoblot using polyclonal or monoclonal antiserum of M.TBH37Rv. 21 days after first vaccination of C57BL-6 mice by all three recombinant eukaryotic expressing vectors, antibody titer for Ag85B reached 1:3200. 21 days after second vaccination, the antibody titer reached 1:102400. The highest antibody levels induced by multivalent vaccines after the second injection were equal to or even greater than the展开更多
Using cDNA representational difference analysis (cDNA RDA) method, we have successfully isolated a gene fragment whose expression was specifically induced by external GAa application. Screening a G2 pea cDNA library u...Using cDNA representational difference analysis (cDNA RDA) method, we have successfully isolated a gene fragment whose expression was specifically induced by external GAa application. Screening a G2 pea cDNA library using this fragment as a probe, we obtained a 2036 bp full-length cDNA. It contains a 1746 bp open reading frame and encodes a protein of 581 amino acids with a theoretical molecular weight of 64 ku. It shares high-level sequenceidentity with AAIR genes from other plant species. This cDNA was cloned into expression vector and recombinant E. coli DH5α cells with remarkable AAIR enzyme activity were obtained.展开更多
Cotton fibers, commonly known as cotton lint, are single-celled trichomes derived from epidermal lay- ers of cotton ovules. Despite of its importance in word trade, the molecular mechanisms of cotton fiber production ...Cotton fibers, commonly known as cotton lint, are single-celled trichomes derived from epidermal lay- ers of cotton ovules. Despite of its importance in word trade, the molecular mechanisms of cotton fiber production is still poorly understood. Through transcriptome profiling, functional genomics, pro- teomics, metabolomics approaches as well as marker-assisted molecular breeding, scientists in China have made significant contributions in cotton research. Here, we briefly summarize major progresses made in Chinese laboratories, and discuss future directions and perspectives relative to the develop- ment of this unique crop plant.展开更多
Based upon analysis of data obtained from theATH1 microarrays, a cDNA that was highly induced afterdrought treatment, was isolated from Arabidopsis seedlings.RT-PCR and Quantitative Real-Time (QRT)-PCR experi-ments sh...Based upon analysis of data obtained from theATH1 microarrays, a cDNA that was highly induced afterdrought treatment, was isolated from Arabidopsis seedlings.RT-PCR and Quantitative Real-Time (QRT)-PCR experi-ments showed that expression level of the gene increasedsignificantly upon drought, UV, abscisic acid, high salinityand salicylic acid treatments. It was classified as a DREBsubfamily member based on multiple sequence alignmentand phylogenetic characterization. Since it encoded a proteinwith a typical ERF/AP2 DNA-binding domain and containeda glutamine-rich region near its N terminus, we named itQRAP2 (for glutamine-rich AP2). Gel retardation assay re-vealed that QRAP2 was able to form a specific complex withthe previously characterized DRE element while did notshow any affinity to the GCC box or the mutant DRE box.When fused to the GAL4 DNA-binding domain, either full-length QRAP2 or its N-terminus functioned effectively as atrans-activator in the yeast one-hybrid assay with its C-ter-minus completely inactive. Our data indicate that QRAP2could be a new member of the AP2/EREBP transcriptionfactor family involved in activation of down-stream targetgenes in response to environmental stress, especially underdrought conditions.展开更多
A mRNA preferentially expressed in cotton fiber was cloned from fiber total RNA of normal upland cotton TM-1 (wild-type) by using RT-PCR and corresponding cDNA (signed as TM-E6) was sequenced. TM-E6 gene had no intron...A mRNA preferentially expressed in cotton fiber was cloned from fiber total RNA of normal upland cotton TM-1 (wild-type) by using RT-PCR and corresponding cDNA (signed as TM-E6) was sequenced. TM-E6 gene had no intron and contained an open reading frame of 771 bp long, and might encode a peptide of 246 amino acids. Other 4 genes, Fl-E6, Li-E6, N-E6 and Bl-E6, which were homologous to TM-E6 gene, were also isolated from 4 fiber mutants of Fiberless Xu-zhou 142, Ligon lintless, Naked seed and Brown lint, respectively. Sequence analysis of each of these mutant genes revealed many variations in structure and nu-cleotide composition of gene when compared with the sequence of TM-E6 gene. (i) There was a changeable repetitive segment in which GGCTCA (Gly-Ser) was repeated 3-5 times between the 82nd and the 93rd codons in different mutant genes. Since the change of Gly-Ser repetitive segment occurred not only in the mutants but also in the wild-type cotton, the repeat frequency might not be associated with the展开更多
Using the barley catalase cDNA as a probe, the effects of salicylic acid (SA) on catalase activity and gene expression have bene studied. The data indicated that higher than 3 0 mmol/L SA inhibited catalase activity t...Using the barley catalase cDNA as a probe, the effects of salicylic acid (SA) on catalase activity and gene expression have bene studied. The data indicated that higher than 3 0 mmol/L SA inhibited catalase activity to a large extent and Northern blot analysis demonstrated that the decrease in catalase activity coincided with a decrease in its mRNA level. The important role of SA in plant systemic acquired resistance (SAR) and its possible mechanism are discussed.展开更多
基金supported by the National Natural Science Foundation of China(90717009)
文摘The biosynthesis and signaling of plant hormones play a critical role in almost all biological processes.It is well-documented that phytohormones cross-talk with each other.Epigenetic mechanisms were suggested to regulate expression of downstream targets in hormone signaling pathways that help implement hormone functions.This new layer of complexities that integrate epigenetic information such as DNA methylation,chromatin remodeling,histone modification,microRNAs and siRNAs with plant hormone signaling and regulations of gene expression,has been gradually revealed.In this short review,the author tries to assemble recent progress to establish a molecular linkage between these two large and momentum research fields and also to help readers digest the literature.
基金This work was supported by the Chinese National High-Tech "863" Project (Grant No. 2001AA213141).
文摘The coding regions of Ag85B MPT-64, and ESAT-6 secreted proteins were cloned initially into the eu-karyotic expression vector pJW4303, then transformed to E. coli Top 10 strain for plasmid DNA extraction and further analysis. Plasmids containing the right insertion were se-quenced to confirm their identity. COS7 cells were trans-fected with a mixture containing serially diluted plasmid DNA encoding three secreted proteins and Lipofectin (Gibco). The supernatants and pellets prepared from various cell lines were run on SDS-PAGE gel and the expression of these proteins in COS7 cells were demonstrated by immunoblot using polyclonal or monoclonal antiserum of M.TBH37Rv. 21 days after first vaccination of C57BL-6 mice by all three recombinant eukaryotic expressing vectors, antibody titer for Ag85B reached 1:3200. 21 days after second vaccination, the antibody titer reached 1:102400. The highest antibody levels induced by multivalent vaccines after the second injection were equal to or even greater than the
基金This work was supported by the National Science Foundation for Distinguished Young Scholars (Grant No. 39725002) and the National "863" High-Tech Project (Grant No. 101-01-01-02).
文摘Using cDNA representational difference analysis (cDNA RDA) method, we have successfully isolated a gene fragment whose expression was specifically induced by external GAa application. Screening a G2 pea cDNA library using this fragment as a probe, we obtained a 2036 bp full-length cDNA. It contains a 1746 bp open reading frame and encodes a protein of 581 amino acids with a theoretical molecular weight of 64 ku. It shares high-level sequenceidentity with AAIR genes from other plant species. This cDNA was cloned into expression vector and recombinant E. coli DH5α cells with remarkable AAIR enzyme activity were obtained.
基金Supported by the National Basic Research Program of China (Grant No. 2004CB117302) Program for Changjiang Scholars and Innovative Research Team in University (Grant No. IRT0432)
文摘Cotton fibers, commonly known as cotton lint, are single-celled trichomes derived from epidermal lay- ers of cotton ovules. Despite of its importance in word trade, the molecular mechanisms of cotton fiber production is still poorly understood. Through transcriptome profiling, functional genomics, pro- teomics, metabolomics approaches as well as marker-assisted molecular breeding, scientists in China have made significant contributions in cotton research. Here, we briefly summarize major progresses made in Chinese laboratories, and discuss future directions and perspectives relative to the develop- ment of this unique crop plant.
基金This work was supported by the National Natural Science Foundation of China(Grant No.30221 120261).
文摘Based upon analysis of data obtained from theATH1 microarrays, a cDNA that was highly induced afterdrought treatment, was isolated from Arabidopsis seedlings.RT-PCR and Quantitative Real-Time (QRT)-PCR experi-ments showed that expression level of the gene increasedsignificantly upon drought, UV, abscisic acid, high salinityand salicylic acid treatments. It was classified as a DREBsubfamily member based on multiple sequence alignmentand phylogenetic characterization. Since it encoded a proteinwith a typical ERF/AP2 DNA-binding domain and containeda glutamine-rich region near its N terminus, we named itQRAP2 (for glutamine-rich AP2). Gel retardation assay re-vealed that QRAP2 was able to form a specific complex withthe previously characterized DRE element while did notshow any affinity to the GCC box or the mutant DRE box.When fused to the GAL4 DNA-binding domain, either full-length QRAP2 or its N-terminus functioned effectively as atrans-activator in the yeast one-hybrid assay with its C-ter-minus completely inactive. Our data indicate that QRAP2could be a new member of the AP2/EREBP transcriptionfactor family involved in activation of down-stream targetgenes in response to environmental stress, especially underdrought conditions.
基金This work was supported by the National Natural Science Foundation of China (Grant Nos. 39770473 and 39830240) and China National Program of Plant Gene Transfer.
文摘A mRNA preferentially expressed in cotton fiber was cloned from fiber total RNA of normal upland cotton TM-1 (wild-type) by using RT-PCR and corresponding cDNA (signed as TM-E6) was sequenced. TM-E6 gene had no intron and contained an open reading frame of 771 bp long, and might encode a peptide of 246 amino acids. Other 4 genes, Fl-E6, Li-E6, N-E6 and Bl-E6, which were homologous to TM-E6 gene, were also isolated from 4 fiber mutants of Fiberless Xu-zhou 142, Ligon lintless, Naked seed and Brown lint, respectively. Sequence analysis of each of these mutant genes revealed many variations in structure and nu-cleotide composition of gene when compared with the sequence of TM-E6 gene. (i) There was a changeable repetitive segment in which GGCTCA (Gly-Ser) was repeated 3-5 times between the 82nd and the 93rd codons in different mutant genes. Since the change of Gly-Ser repetitive segment occurred not only in the mutants but also in the wild-type cotton, the repeat frequency might not be associated with the
文摘Using the barley catalase cDNA as a probe, the effects of salicylic acid (SA) on catalase activity and gene expression have bene studied. The data indicated that higher than 3 0 mmol/L SA inhibited catalase activity to a large extent and Northern blot analysis demonstrated that the decrease in catalase activity coincided with a decrease in its mRNA level. The important role of SA in plant systemic acquired resistance (SAR) and its possible mechanism are discussed.
