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适用于流式细胞术的三角褐指藻核DNA染色方法 被引量:2
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作者 张志霞 高莉 +3 位作者 刘倩倩 曹素娟 兰利琼 卿人韦 《应用与环境生物学报》 CAS CSCD 北大核心 2018年第6期1270-1274,共5页
为制备可用于配置有488 nm激发器的流式细胞仪检测核DNA含量的三角褐指藻细胞,取对数生长期的三角褐指藻细胞,分别采用70%乙醇、1%戊二醛、1%甲醛溶液进行细胞固定,经RNase A消化后的样品用碘化丙啶(PI)染色,用SYBR-green I对未经RNase ... 为制备可用于配置有488 nm激发器的流式细胞仪检测核DNA含量的三角褐指藻细胞,取对数生长期的三角褐指藻细胞,分别采用70%乙醇、1%戊二醛、1%甲醛溶液进行细胞固定,经RNase A消化后的样品用碘化丙啶(PI)染色,用SYBR-green I对未经RNase A消化的细胞染色,在荧光显微镜下观察细胞染色情况,用流式细胞仪检测细胞核DNA含量.结果显示:在荧光显微镜下观察到,3种不同固定液固定的细胞均表现为SYBR-green I只对藻细胞核染色,PI对于藻的细胞核与细胞质均有着色.流式结果的DNA直方图显示,SYBR-green I染色的细胞的DNA直方图有明显的G1与G2/M峰,拟合度(RCS)在3左右,变异系数(CV)在8.6%左右.而PI染色细胞的DNA直方图表现为变异系数值偏大,拟合度较低,没有明显的G1与G2/M峰.本研究表明,SYBR-green I是一种无需对细胞进行RNase A处理,可应用于配置有488 nm激光器流式细胞仪的核酸染料,可为研究在整个细胞周期的调控机制奠定基础,同时也为其他浮游植物细胞核的染色提供参考. 展开更多
关键词 三角褐指藻 流式细胞 碘化丙啶(PI) SYBR-green I 细胞核dna染色
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Fluorescent vital staining of plant sexual cell nuclei with DNA-specific fluorochromes and its application in gametoplast fusion 被引量:2
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作者 YANGHONGYUAN XINLIWU 《Cell Research》 SCIE CAS CSCD 1993年第2期121-130,共10页
DNA-binding fluorochromes are often used for vital staining of plant cell nuclei. However, it is not always sure whether the cells after staining still remain in living state. We chose several criteria to estimate the... DNA-binding fluorochromes are often used for vital staining of plant cell nuclei. However, it is not always sure whether the cells after staining still remain in living state. We chose several criteria to estimate the validity of real vital staining for sexual cell nuclei. These were: the cytoplasmic streaming in pollen tubes whose nuclei were stained, the simultaneous visualization of fluo-rochromatic reaction and nucleus staining in isolated generative cells, and the capability of isolated, prestained generative or sperm cells to fuse with other protoplasts. The results confirmed that 4',6-diamidino-2-phenylindole (DAPI), Hoechst 33258 and mithramycin could be used as real vital stains, though their efficiency varied from case to case; among them DAPI showed best effect. The fluorescent vital staining technique offered a useful means fori-dentification and selection of heterokaryons in gametoplast manipulation studies. 展开更多
关键词 fluorescent vital staining dna-specific fluorochrome generative cell sperm cell gametoplast fusion.
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Recruitment and reinforcement: maintaining epigenetic silencing 被引量:6
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作者 Chengzhi Wang Bing Zhu Jun Xiong 《Science China(Life Sciences)》 SCIE CAS CSCD 2018年第5期515-522,共8页
Cells need to appropriately balance transcriptional stability and adaptability in order to maintain their identities while responding robustly to various stimuli. Eukaryotic cells use an elegant "epigenetic"... Cells need to appropriately balance transcriptional stability and adaptability in order to maintain their identities while responding robustly to various stimuli. Eukaryotic cells use an elegant "epigenetic" system to achieve this functionality. "Epigenetics" is referred to as heritable information beyond the DNA sequence, including histone and DNA modifications, nc RNAs and other chromatin-related components. Here, we review the mechanisms of the epigenetic inheritance of a repressive chromatin state,with an emphasis on recent progress in the field. We emphasize that(i) epigenetic information is inherited in a relatively stable but imprecise fashion;(ii) multiple cis and trans factors are involved in the maintenance of epigenetic information during mitosis; and(iii) the maintenance of a repressive epigenetic state requires both recruitment and self-reinforcement mechanisms.These mechanisms crosstalk with each other and form interconnected feedback loops to shape a stable epigenetic system while maintaining certain degrees of flexibility. 展开更多
关键词 heterochromatin heterochromatin maintenance polycomb H3K27me3 H3K9me2/3
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