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Development of oligonucleotide probes for FISH karyotyping in Haynaldia villosa,a wild relative of common wheat

The Crop Journal 2020年第4期8卷 676-681页

作者： Jia Lei Jiawen Zhou Haojie Sun Wentao Wan Jin Xiao Chunxia Yuan Miroslava Karafiátová Jaroslav Dolezel Haiyan Wang Xiue Wang State Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University/JCIC-MCPNanjing 210095JiangsuChina Institute of Experimental Botany Czech Academy of SciencesCentre of the Region Hana for Biotechnological and Agricultural ResearchŠlechtitelů3177900 OlomoucCzech Republic

Haynaldia villosa is a wild relative of wheat and a valuable gene resource for wheat *** to the limited number of probes available for fluorescence in situ hybridization(FISH),the resolution at which the karyotype of *** can be characterized is poor,hampering accurate characterization of small segmental alien *** designed ten oligonucleotide probes using tandem repeats in DNA sequences derived from the short arm of *** chromosome 6 V(6 VS).FISH with seven of them resulted in clear signals on *** *** these,we constructed FISH karyotypes for *** using oligo-6 VS-1 and oligo-6 VS-35 oligonucleotides and characterized the distribution of the two probes in five different *** *** new FISH probes can efficiently characterize *** introgressions into wheat.

关键词： Chromosome identification Haynaldia villosa oligonucleotide probes Tandem DNA repeats

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Establishment and characterization of a rat pancreatic stellate cell line by spontaneous immortalization

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World Journal of Gastroenterology 2003年第12期9卷 2751-2758页

作者： Atsushi Masamune Masahiro Satoh Kazuhiro Kikuta Noriaki Suzuki Tooru Shimosegawa Division of Gastroenterology Tohoku University Graduate School of MedicineSendaiJapan A.M.and M.S.equally contributed to this work

AIM: Activated pancreatic stellate cells (PSCs) have been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line may therefore be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish a cell line of rat PSCs by spontaneous ***: PSCs were isolated from the pancreas of male Wistar rats, and conventional subcultivation was performed repeatedly. Telomerase activity was measured using the telomere repeat amplification protocol. Activation of transcription factors was assessed by electrophoretic mobility shift *** of mitogen-activated protein (MAP) kinases was examined by Western blotting using anti-phosphospecific antibodies. Expression of cytokine-induced neutrophil chemoattractant-1 was determined by enzyme ***: Conventional subcultivation yielded actively growing cells. One clone was obtained after limiting dilution,and designated as SIPS. This cell line has been passaged repeatedly more than 2 years, and is thus likely *** cells retained morphological characteristics of primary,culture-activated PSCs. SIPS expressed α-smooth muscle actin, glial acidic fibrillary protein, vimentin, desmin, type Ⅰ collagen, fibronectin, and prolyl hydroxylases. Telomerase activity and p53 expression were negative. Proliferation of SIPS cells was serum-dependent, and stimulated with platelet-derived growth factor-BB through the activation of extracellular signal-regulated kinase. Interleukin-1β activated nuclear factor-κB, activator protein-1, and MAP ***-1β induced cytokine-induced neutrophil chemoattractant-1 expression through the activation of nuclear factor-κB and MAP ***: SIPS cells can be useful for in vitro studies of cell biology and signal transduction of PSCs.

关键词： Animals Base Sequence Cell Culture Techniques Cell Line, Transformed Cells, Cultured Cystic Fibrosis Cytoskeletal Proteins Extracellular Matrix Proteins Immunohistochemistry NF-kappa B oligonucleotide probes Pancreas Rats Research Support, Non-U.S. Gov't Transcription Factor AP-1

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Detection of two pathogenic marine ciliates Ancistrum haliotis and A. crassum(Ciliophora: Scuticociliatia) by fluorescence in situ hybridization

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Acta Oceanologica Sinica 2020年第12期39卷 90-94页

作者： Zifeng Zhan Kuidong Xu Laboratory of Marine Organism Taxonomy and Phylogeny Institute of OceanologyChinese Academy of SciencesQingdao 266071China Laboratory for Marine Biology and Biotechnology Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China Center for Ocean Mega-Science Chinese Academy of SciencesQingdao 266071China University of Chinese Academy of Sciences Beijing 100049China

The scuticociliatid ciliates Ancistrum haliotis and *** are parasites that may cause high mortality in the cultured abalone Haliotis *** the bivalve Ruditapes *** identification with silver staining methods is hampered by their morphological similarities to closely related species and the complicated procedures of morphological *** designed two SSU rRNA-targeted oligonucleotide probes labeled with a fluorochrome,and optimized the fluorescence in situ hybridization(FISH)protocols for identification of *** and ***,*** assays resulted in a clear identification by strong fluorescence signals from the oligonucleotide *** method can be used for quick and accurate quantitative analysis of *** and *** infections on host molluscs.

关键词： oligonucleotide probes molecular methods scuticociliates SSU rRNA abalone mollusc

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Fast species identification of mycobacterium by rpoB gene chip technology

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Journal of Life Sciences 2009年第5期3卷 43-46页

作者： LI Hong-min FAN Bo WANG Wei AN Hui-ru MIAO Qing Tuberculosis Research Laboratory The Second Clinical Department of the PLA General Hospital Beijing 100091 China

Based on rpoB gene micro array as target gene, we are going to use gene chip technology to test 24 mycobacterium standard specimens, 8 non-mycobacterium specimens and 86 mycobacterium clinical isolated specimens. As a result, after mycobacterium and non-mycobacterium standard specimens were duplicated by PCR, mycobacterium standard specimens reproduced 360bp DNA fragments; on the other hand, non-mycobacterium specimens did not reproduce any fragments except for hemolytic streptococcus and corynebacterium pseudodiphtheriticum which had the same results as mycobacterium standard specimens. Sensitive test is able to detect lpg tuberculosis mycobacterium DNA. The probe test showed that, among 21 oligonucleotide probes, probe-M. fortuitum and M. marinum were cross-hybrid; the other probes were specific. We used the new method to identify 126 mycobacterium clinical isolated specimens. The test results of this new method matched with conventional method. In conclusion, compared to the traditional method, the use of rpob gene chip technology to identify mycobacterium species will be faster, more accurate and higher value in application.

关键词： mycobacterium rpoB gene chip technology oligonucleotide probes

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Rapid and Accurate Detection of Plant miRNAs by Liquid Northern Hybridization

Rapid and Accurate Detection of Plant miRNAs by Liquid North...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者：王小粟童永鳌王胜华四川大学生命科学学院

Northern blot analysis is a powerful research tool for discovery,validation and expression of genes,and is currently widely used to detect microRNA(miRNA) ***,the traditional Northern blot procedure,which is based on a support membrane,is overly elaborate and time-consuming, although it is unsurpassed in accuracy for determining the sizes and amounts of multiple small RNAs sharing high sequence *** we present an alternative method derived from plant miRNAs,liquid Northern hybridization,using fluorescently labeled oligonucleotide probes and characterized by simple and specific miRNA determination and *** entire detection process is completed within a few hours,and multiple miRNAs can be simultaneously detected in a single experiment.

关键词： miRNA liquid Northern hybridization Oryza sativa small RNA oligonucleotide probes

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