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In order to investigate the molecular mechanism of silk/threonine protein kinase (STK)-mediated blue light response in the algal Chlamydomonas reinhardtii, phenotype identification and transcriptome analysis were conducted for C. reinhardtii STK mutant strain crstk11 (with an AphvIII box reverse insertion in stk11 gene coding region) under blue light stress. Phenotypic examination showed that under normal light (white light), there was a slight difference in growth and pigment contents between the wild-type strain CC5325 and the mutant strain crstk11. Blue light inhibited the growth and chlorophyll synthesis in crstk11 cells, but significantly promoted the accumulation of carotenoids in crstk11. Transcriptome analysis showed that 860 differential expression genes (DEG) (559 up-regulated and 301 down-regulated) were detected in mutant (STK4) vs. wild type (WT4) upon treatment under high intensity blue light for 4 days. After being treated under high intensity blue light for 8 days, a total of 1 088 DEGs (468 upregulated and 620 downregulated) were obtained in STK8 vs. WT8. KEGG enrichment analysis revealed that compared to CC5325, the crstk11 blue light responsive genes were mainly involved in catalytic activity of intracellular photosynthesis, carbon metabolism, and pigment synthesis. Among them, upregulated genes included psaA, psaB, and psaC, psbA, psbB, psbC, psbD, psbH, and L, petA, petB, and petD, as well as genes encoding ATP synthase α, β and c subunits. Downregulated genes included petF and petJ. The present study uncovered that the protein kinase CrSTK11 of C. reinhardtii may participate in the blue light response of algal cells by mediating photosynthesis as well as pigment and carbon metabolism, providing new knowledge for in-depth analysis of the mechanism of light stress resistance in the algae.
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Chlamydomonas reinhardtii/genética , Fotosíntesis/genética , Plantas/metabolismo , Proteínas Quinasas , Treonina/metabolismo , Carbono/metabolismo , Serina/metabolismoRESUMEN
Objective: To investigate the molecular mechanism of inhibitory effect of Scutellaria baicalensis to uropathogenic Escherichia coli (UPEC). Methods: RNA-seq was used to analyze transcriptome of UPEC NB8 effected by water decoction of S. baicalensis. S. baicalensis group and negative control group were set up. In S. baicalensis group, UPEC NB8 was disposed by water decoction of 10 times of MIC of S. baicalensis (62.5 mg/mL) for 30 min. While in negative control group, UPEC NB8 was disposed by same amount of normal saline. Then, total RNA of UPEC NB8 was extracted, while rRNA was removed, cDNA was reverse transcriptional synthesized. Transcriptome sequencing was sequenced on HiSeq2000, transcriptome data were analyzed by BIGpre, Tophat, and Cufflinks. Expression profile was analyzed for differential expression, GO, COG functional enrichment analysis, and KEGG metabolic pathway analysis. Results: A total of 1665 differentially expressed genes existed between S. baicalensis group and negative control group. Among them, 1169 genes were up-regulated genes, while 496 genes were down-regulated genes. Under the effect of water decoction of S. baicalensis, genes of NB8 down-regulated in glycolysis, Krebs cycle, biosynthesis of fatty acids, and ribosomal proteins, while genes of NB8 up-regulated chemotaxis and assembly path of flagella, and transportation protein ABC pathway. Conclusion: The molecular mechanism of the inhibitory effect of S. baicalensis to UPEC is explained. Targets of S. baicalensis to UPEC are glycolysis, Kreb's cycle, biosynthesis of fatty acids, and translation of proteins. Besides, chemotaxis, the assembly path of flagella, and transportation protein ABC also play a key role in alarm reaction of S. baicalensis.
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OBJECTIVE: To investigate the gene differential expression in Pesudostellariae Radix from different habitats, isolate and evaluate associated differential genes. METHODS: cDNA-AFLP technique was applied to analyze the differential expression genes of Pesudostellariae Radix from four different habitats. RESULTS: Six primer pairs were selected and amplified. Thirty-four differentially expressed trivially distributed file system (TDFs) were obtained from Pesudostellariae Radix samples from different habitats. Then these TDFs were cloned and sequenced, and the nucleotide sequences of 21 TDFs were obtained. By BLASTX analysis, 15 of them were found to have homologous sequences in the databases, among which seven TDFs had known function. These proteins were mainly involved in the growth and development of plants and played a role in the metabolisms of defending diseases and insect pests, and improving the ability of plants to resist abiotic stress. CONCLUSION: This research evaluates differential expression genes in Pesudostellariae Radix from different habitats, providing the basic information for revealing the molecular mechanism of the property formation of Pesudostellariae Radix.
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Objective To screen for Kaposi sarcoma (KS)-related genes. Methods Tissue samples were obtained from the lesion and normal skin of a patient with KS in Xinjiang Uygur Autonomous Region,and total RNA was extracted from these samples and reverse transcribed into cDNA. Real-time fluorescent quantitative reverse transcription PCR (RT-qPCR) was performed to determine the expression of K8.1, K2 and ORF50 in these samples. The cDNA was labeled with fluorescein and hybridized to a human 35K genome array containing 25 100 genes. Subsequently, the signal images were scanned by a laser scanner and acquired images were analyzed by software. Results RT-qPCR revealed the mRNA expression of K8.1, K2 and ORF50 in the KS tissues but not in the normal skin tissues, indicating that there was no crossed contamination in these specimens. Among the 25 100 genes, 1313 genes were identified to be differentially expressed between KS and normal skin tissues, including 756 up-regulated genes and 557 down-regulated genes. These differentially expressed genes, such as myeloid cell leukemia-1 gene (MCI-1), annexins (ANX) and serine proteinase inhibitor Kazal type 5 (SPINK5), were associated with apoptosis, angiogenesis, cell signaling, protein processing, cell cycle regulation, and so on. Conclusion The differentially expressed genes such as MCI-1 and SPINK5 may be associated with the development of KS.
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30% was considered sensitive and the rate ≤30% was considered resistant;and the 6 specimens were divided into 2 group according to the above standard.cDNA microassay combined with clustering analysis was used to screen for resistance-related genes.Semi-quantitative RT-PCR was used for verification of HDAC1 gene expression.Results:Three of the 6 glioma specimens belonged to the drug resistance group and the other 3 to the drug sensitive group.cDNA microarray analysis combined with cluster analysis screened out 21 genes,with 6 up-regulated and 15 down-regulated.High expression of gene HDAC1 was noticed in all the 6 specimens by semi-quantitative RT-PCR,and the trend was similar to that by microassay.Conclusion:The primary drug resistance of glioma may be associated with the 21 genes screened by cDNA microarray;the detailed mechanisms for drug controlling still need to discussed in the future.