mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the ...mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the maternal RNAs stored in egg. These mRNAs are being degraded as the development proceeds. In some animals, such as fish and amphibian, new transcripts do not appear until the midblastula stage (midblastula transition, MBT). In other animals, for example in mouse, the zygotic genes are expressed during very early stages of development. The mRNA programmed synthesis and degradation during embryonic development controls the cell differentiation, germlayer formation and pattern formation. All these mRNA changes could be displayed side by side as cDNA band differences by mRNA differential display and the genes corresponding to these differential mRNAs could thus be obtained.展开更多
Flatfish or flounder moves one eye to change body proportion into vertebral asymmetry during metamorphosis, during which some become sinistral while others dextral. However, the mechanism behinds the eye-position has ...Flatfish or flounder moves one eye to change body proportion into vertebral asymmetry during metamorphosis, during which some become sinistral while others dextral. However, the mechanism behinds the eye-position has not been well understood. In this research, hybrids between Japanese flounder(♀) and stone flounder (♂) show mixed eye-location in both dextral type and sinistral type, and thus become good samples for studying the eye-migration. mRNAs from pro-metamorphosis sinistral and dextral hybrids larvae were screened with classical differential display RT-PCR (DD-RT-PCR) and representational difference analysis of cDNA (cDNA-RDA); 30 and 47 putative fragments were isolated, respectively. The cDNA fragments of creatine kinase and trypsinogen 2 precursor genes isolated by cDNA-RDA exhibited eye-position expression patterns during metamorphosis. However, none of the fragments was proved to be related to flatfishes’ eye-position specifically. Therefore, further studies and more sensitive gene isolated methods are needed to solve the problems.展开更多
Objective： The aim of the study was to establish an arsenic trioxide （ATO）-resistant cell line of human gallbladder carcinoma, GBC-SD/ATO, and to analyze the differential expressions of its apoptosis-associated gen...Objective： The aim of the study was to establish an arsenic trioxide （ATO）-resistant cell line of human gallbladder carcinoma, GBC-SD/ATO, and to analyze the differential expressions of its apoptosis-associated genes, so as to investigate a correlation between ATO induced resistance of gallbladder carcinoma and expressions of apoptosis associated genes. Methods： The resistant cell line was obtained in vitro by culture of human gallbladder carcinoma cell line GBC-SD with increasingly stepwise concentrations of ATO. The sensitivities of GBC-SD cells and GBC-SD/ATO cells to ATO were determined by MTT assay respectively, cDNA microarray containing 458 apoptosis-related human genes was used to compare the gene expression profiles of GBC-SD/ATO cells and corresponding sensitive cell line GBC-SD. Results： GBC-SD/ATO cell line was established successfully after 8 months of exposure to increasing concentrations of ATO. Compared with the parental cell line, GBC-SD/ATO was 13.6 times more resistant to ATO. Of the 458 apoptosis-related genes, 17 genes were detected having 〉 2-fold difference of expression between the GBC-SD/ATO and GBC-SD cells, with 6 genes up-regulated and 11 genes down-regulated in GBC-SD/ATO cells. Conclusion： The 17 genes invoJved in the apoptosis pathway might be relevant to the resistance of GBC-SD/ATO cells to ATO, suggesting that the modulation of expression of apoptosis-related genes may be a main mechanism of acquired resistance in GBC-SD/ATO.展开更多
Panax ginseng C. A. Meyer is an important traditional herb in eastern Asia. It contains ginsenosides, which are primary bioactive compounds with medicinal properties. Although ginseng has been cultivated since at leas...Panax ginseng C. A. Meyer is an important traditional herb in eastern Asia. It contains ginsenosides, which are primary bioactive compounds with medicinal properties. Although ginseng has been cultivated since at least the Ming dynasty to increase production, cultivated ginseng has lower quantities of ginsenosides and lower disease resistance than ginseng grown under natural conditions. We extracted root RNA from six varieties of fifth-year P. ginseng cultivars representing four different growth conditions, and performed Illumina paired-end sequencing. In total, 163,165,706 raw reads were obtained and used to generate a de novo transcriptome that consisted of 151,763 contigs(76,336 unigenes), of which 100,648 contigs(66.3%) were successfully annotated. Differential expression analysis revealed that most differentially expressed genes(DEGs) were upregulated(246 out of 258, 95.3%) in ginseng grown under natural conditions compared with that grown under artificial conditions. These DEGs were enriched in gene ontology(GO) terms including response to stimuli and localization. In particular, some key ginsenoside biosynthesis-related genes, including HMG-Co A synthase(HMGS), mevalonate kinase(MVK), and squalene epoxidase(SE), were upregulated in wild-grown ginseng. Moreover, a high proportion of disease resistance-related genes were upregulated in wild-grown ginseng. This study is the first transcriptome analysis to compare wild-grown and cultivated ginseng, and identifies genes that may produce higher ginsenoside content and better disease resistance in the wild; these genes may have the potential to improve cultivated ginseng grown in artificial environments.展开更多
文摘mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the maternal RNAs stored in egg. These mRNAs are being degraded as the development proceeds. In some animals, such as fish and amphibian, new transcripts do not appear until the midblastula stage (midblastula transition, MBT). In other animals, for example in mouse, the zygotic genes are expressed during very early stages of development. The mRNA programmed synthesis and degradation during embryonic development controls the cell differentiation, germlayer formation and pattern formation. All these mRNA changes could be displayed side by side as cDNA band differences by mRNA differential display and the genes corresponding to these differential mRNAs could thus be obtained.
基金Supported by National Natural Science Foundation of China (No. 30600455)the National Basic Research Program of China (973 Program, No.2004CB117402)
文摘Flatfish or flounder moves one eye to change body proportion into vertebral asymmetry during metamorphosis, during which some become sinistral while others dextral. However, the mechanism behinds the eye-position has not been well understood. In this research, hybrids between Japanese flounder(♀) and stone flounder (♂) show mixed eye-location in both dextral type and sinistral type, and thus become good samples for studying the eye-migration. mRNAs from pro-metamorphosis sinistral and dextral hybrids larvae were screened with classical differential display RT-PCR (DD-RT-PCR) and representational difference analysis of cDNA (cDNA-RDA); 30 and 47 putative fragments were isolated, respectively. The cDNA fragments of creatine kinase and trypsinogen 2 precursor genes isolated by cDNA-RDA exhibited eye-position expression patterns during metamorphosis. However, none of the fragments was proved to be related to flatfishes’ eye-position specifically. Therefore, further studies and more sensitive gene isolated methods are needed to solve the problems.
基金Supported by the grants from the Research Fund of the Educational Department of Zhejiang Province (No 20070609)Natural Science Foundation of Zhejiang Province (No Y206860)Natural Science Foundation of Zhejiang Province (No Y207802)
文摘Objective： The aim of the study was to establish an arsenic trioxide （ATO）-resistant cell line of human gallbladder carcinoma, GBC-SD/ATO, and to analyze the differential expressions of its apoptosis-associated genes, so as to investigate a correlation between ATO induced resistance of gallbladder carcinoma and expressions of apoptosis associated genes. Methods： The resistant cell line was obtained in vitro by culture of human gallbladder carcinoma cell line GBC-SD with increasingly stepwise concentrations of ATO. The sensitivities of GBC-SD cells and GBC-SD/ATO cells to ATO were determined by MTT assay respectively, cDNA microarray containing 458 apoptosis-related human genes was used to compare the gene expression profiles of GBC-SD/ATO cells and corresponding sensitive cell line GBC-SD. Results： GBC-SD/ATO cell line was established successfully after 8 months of exposure to increasing concentrations of ATO. Compared with the parental cell line, GBC-SD/ATO was 13.6 times more resistant to ATO. Of the 458 apoptosis-related genes, 17 genes were detected having 〉 2-fold difference of expression between the GBC-SD/ATO and GBC-SD cells, with 6 genes up-regulated and 11 genes down-regulated in GBC-SD/ATO cells. Conclusion： The 17 genes invoJved in the apoptosis pathway might be relevant to the resistance of GBC-SD/ATO cells to ATO, suggesting that the modulation of expression of apoptosis-related genes may be a main mechanism of acquired resistance in GBC-SD/ATO.
基金supported by the International Science and Technology Cooperation of China(2011DFA32730)
文摘Panax ginseng C. A. Meyer is an important traditional herb in eastern Asia. It contains ginsenosides, which are primary bioactive compounds with medicinal properties. Although ginseng has been cultivated since at least the Ming dynasty to increase production, cultivated ginseng has lower quantities of ginsenosides and lower disease resistance than ginseng grown under natural conditions. We extracted root RNA from six varieties of fifth-year P. ginseng cultivars representing four different growth conditions, and performed Illumina paired-end sequencing. In total, 163,165,706 raw reads were obtained and used to generate a de novo transcriptome that consisted of 151,763 contigs(76,336 unigenes), of which 100,648 contigs(66.3%) were successfully annotated. Differential expression analysis revealed that most differentially expressed genes(DEGs) were upregulated(246 out of 258, 95.3%) in ginseng grown under natural conditions compared with that grown under artificial conditions. These DEGs were enriched in gene ontology(GO) terms including response to stimuli and localization. In particular, some key ginsenoside biosynthesis-related genes, including HMG-Co A synthase(HMGS), mevalonate kinase(MVK), and squalene epoxidase(SE), were upregulated in wild-grown ginseng. Moreover, a high proportion of disease resistance-related genes were upregulated in wild-grown ginseng. This study is the first transcriptome analysis to compare wild-grown and cultivated ginseng, and identifies genes that may produce higher ginsenoside content and better disease resistance in the wild; these genes may have the potential to improve cultivated ginseng grown in artificial environments.
