A protocol for expression of foreign genes in chloroplasts
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Chloroplasts are plant-specific organelles that perform photosynthesis and responsible for the world's primary productivity. Using light energy, chloroplasts produce many important products, including starch, amino acids, lipids, pigments and various secondary products. Therefore, chloroplasts are essential to the lives of all plants and animals alike. Chloroplast transformation is a unique technology to produce huge amount of valuable materials in chloroplasts using photosynthetic energy. In order to control chloroplasts at will, we need more information on molecular basis of chloroplast gene expression and communication between the chloroplast and nucleus. Chloroplast sigma factors are key regulators of the chloroplast gene expression and chloroplast differentiation. This review summarized recent findings on roles of chloroplast sigma factors in the chloroplast differentiation and environmental responses.
Organelle
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Objective: To screen the differential expression genes of Kanglaite Injection in treating cancer cachexia. Methods: mRNA was extracted from the blood cells of T739 animal model of C.C., hybridizated respectively on 20S gene chip. Analysis discuss on differential expression genes was carried out. Results: 5 differential expression genes were obtained. Among these genes, 4 genes were up-regulated and 1 gene was down-regulated. Most of these genes were related with immunity and metabolism of tumor. Conclusion: cDNA microarray for analysis of gene expression patterns is a powerful method to identify associated genes of Kanglaite.
Cancer Cachexia
Gene chip analysis
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ABSTRACT It is not understood what evolutionary factors drive some genes to be expressed at a higher level than others. Here, we hypothesized that a gene’s function plays an important role in setting expression level. First, we established that each S. cerevisiae gene is maintained at a specific expression level by analyzing RNA-seq data from multiple studies. Next, we found that mRNA and protein levels were maintained for the orthologous genes in S. pombe , showing that gene function, conserved in orthologs, is important in setting expression level. To further explore the role of gene function in setting expression level, we analyzed mRNA and protein levels of S. cerevisiae genes within gene ontology (GO) categories. The GO framework systematically defines gene function based on experimental evidence. We found that several GO categories contain genes with statistically significant expression extremes; for example, genes involved in translation or energy production are highly expressed while genes involved in chromosomal activities, such as replication and transcription, are weakly expressed. Finally, we were able to predict expression levels using GO information alone. We created and optimized a linear equation that predicted a gene’s expression based on the gene’s membership in 161 GO categories. The greater number of GO categories with which a gene is associated, the more accurately expression could be predicted. Taken together, our analysis systematically demonstrates that gene function is an important determinant of expression level.
Pair-rule gene
Transcription
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Evolutionary rates provide important information about the pattern and mechanism of evolution. Although the rate of gene sequence evolution has been well studied, the rate of gene expression evolution is poorly understood. In particular, it is unclear whether the gene expression level and tissue specificity influence the divergence of expression profiles between orthologous genes. Here we address this question using a microarray data set comprising the expression signals of 10,607 pairs of orthologous human and mouse genes from over 60 tissues per species. We show that the level of gene expression and the degree of tissue specificity are generally conserved between the human and mouse orthologs. The rate of gene expression profile change during evolution is negatively correlated with the level of gene expression, measured by either the average or the highest level among all tissues examined. This is analogous to the observation that the rate of gene (or protein) sequence evolution is negatively correlated with the gene expression level. The impacts of the degree of tissue specificity on the evolutionary rate of gene sequence and that of expression profile, however, are opposite. Highly tissue-specific genes tend to evolve rapidly at the gene sequence level but slowly at the expression profile level. Thus, different forces and selective constraints must underlie the evolution of gene sequence and that of gene expression.
Molecular evolution
Sequence (biology)
Rate of evolution
Divergence (linguistics)
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To study the genes differentially expressed in the liver of Kkay diabetic and normal mice by genomic-scale gene expression analysis.cDNA microarray chips containing 8,192 cDNAs were used to explore the gene expression pattern of Kkay mouse liver.One hundred and fifty-four genes were screened out, including 68 complete cDNAs and expressed sequence tags, and among them 40 genes were up-regulated and 114 genes were down-regulated respectively.Most of the gene expression analysis results were consistent with previous study, and the gene expression pattern of Kkay mouse based on cDNA microarray could be used for high-throughout screening out the genes associated with type 2 diabetes.
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The chloroplast numbers,morphologies and ultra-structures in the leaves of a xantha wheat mutants etiolating naturally and its parent(Xinong1718) were comparatively examined by transmission electron microscopy.It was shown that the mutant plants with three etiolating degrees did not obviously differ in chloroplast distribution,number,shape and size from the mutant parent;(2) Yellowish-green plants whose chlorophyll content was 58% of the wild type did not obviously differed in chloroplast ultra-structure from the parent,but their stroma-thylakoids and granum-thylakoids were highly differentiated and their granum and grana lamellae were relatively plentiful;(3) The chlorophyll contents of the golden and greenish-yellow plants were separately 17% and 24% of that of the wild type and their chloroplast ultra-structures obviously differed from that of the parent and the chloroplasts of the mutant plants had clear developmental defect,the chloroplasts of golden plants containing no grana,obviously visible stroma lamellae,starch grains and plentiful osmiophilic granules and those of greenish-yellow mutant plants containing grana whose number was obviously lower than that in the chloroplasts of the parent and that had a fewer layers of stroma lamellae,and highly developed granum-thylakoids.The results indicted that the change in the chloroplast ultra-structure of the xantha wheat mutant resulted from lowered chlorophyll content and it was supposed that the etiolating mutation resulted from chlorophyll synthesis blockade.
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The amount of total chlorophyll, chlorophylls a and b as well as the ratio of a to b decreased in chloroplasts isolated from Quercus rotundifolia leaves, kept for 17 d in a solution of 35.5 μM evernic acid in 1 mM Na HCO3, when compared with the chloroplasts of control leaves (kept in NaHCO3). The chloroplasts in the spongy parenchyma were smaller and the amount of starch and plastoglobuli lower. The number of grana per chloroplast section, the number of thylakoids per grana and the height of grana stacks were also less in the chloroplasts of leaves treated with evernic acid. Quantitative ultrastructural differences were determined by means of electron microscopy and image analysis techniques.
Parenchyma
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Objective To analyze the differential gene expression profiling of liver in rats subjected to hemorrhagic shock(HS) and sham hemorrhage shock(SHAM) by gene chip technology, thus to evaluate the possible molecular pathogenesis of HS. Method 20 male Wistar rats were randomly divided into a SHAM group and a HS group, with 10 rats in each group. Hepatic gene expression profiles were detected by oligonucleotide microarrays of 5705 mouse genes in two groups for three times. Genes with ratio(R) > 2 were identified as up-regulated and R < 0.5 were identified as down-regulated. Biological function of differentially expressed genes was analyzed and 9 genes were selected to undergo semi-quantitative RT-PCR. Results Among the total 5705 probes detected,86 genes showed differential expression in HS group comparison with SHAM group. The expression levels of 72 genes were up-regulated while those of 14 genes were down-regulated significantly. Differentially expressed genes were classified according to their biological function: transport genes, transcription regulator genes, signaling genes, response to stress genes, metabolic genes, development genes and cell adhesion genes. Conclusions cDNA microarray is an efficient and high-throughout method to survey gene expression profiles in HS.The variation of those gene expressions might be a potential pathogenic mechanism for HS that may offer a novel target for further study of therapeutic strategies of HS.
Key words:
Hemorrhagic shock; DNA chip; Gene expression; liver
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엽록체 형질전환된 식물체를 얻으려면 먼저 세포수준에서 모든 엽록체가 형질전환되어야 하는데, 세포내에는 많은 수의 엽록체가 존재하므로, 엽록체 형질전환 벡터가 전이되어 형질전환된 엽록체는 선발배지에서 선택적으로 분열을 계속하고 형질전환되지 않은 엽록체들은 분열을 하지 못하게 되어, 결국 해당 세포내의 모든 엽록체가 형질전환된 상태에 이르게 된다. 따라서 만일 해당 세포내에 엽록체의 수가 적으면 그만큼 효율적으로 엽록체 형질전환을 할 수 있을 것이다. 본 연구에서는 담배의 FtsZ 유전자를 핵형질전환법으로 과잉 발현시킴으로써 엽록체의 분열이 저해되어 엽육세포내에 거대한 엽록체 3-5개를 가진 담배식물체의 엽육조직을 이용하여, 엽록체 형질전환을 한 결과, 엽록체 형질전환 빈도가 약 40% 증가되었다. In the chloroplast transformation process, a chloroplast containing transformed chloroplast genome copies should be selected over wild-type chloroplasts on selection medium. It is more effective for a cell to become homoplasmic if the cell contains smaller number of chloroplasts. Therefore, to reduce the number of chloroplasts in mesophyll cells in tobacco, we overexpressed FtsZ to generate transgenic plants, of which mesophyll cell contained a few enlarged chloroplasts contrast to a wild-type mesophyll cell containing approximately 100 chloroplasts. It was demonstrated that transgenic leaf tissues comprising cells with a few enlarged chloroplasts gave rise to approximately 40% higher frequency of chloroplast-transformed adventitious shoots.
FtsZ
Nuclear gene
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