说明酸谈实验室.pdfVIP

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  • 2025-05-09 发布于北京
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染色质密度(ChromatinDensity)

Electronmicroscopyimageshowingthe

classicaldichotomybetweeneuchromatinand

heterochromatininahumanleucocyte.Mostof

heterochromatin(blackarrows)isintheperipheryof

thenucleus,forthemostparttightlyattachedtothe

lamina,whileeuchromatin(spacepointedbythe

whitearrows)isfillingmostofthenucleusspace,

withouttouchinglamina.Imagecourtesy:Wioleta

Dudka.

KIRTIPRAKASH.THEPERIODICANDDYNAMIC

STRUCTUREOFCHROMATIN.

细胞核中有常染色质和异染色质

常染色质上的基因就可能处于转录活跃状态

ONDinducescompactionofchromatintomechanisticallyreducetranscription.HL-1cells

weresubjectedtobothanti-H3K14immunostainingandcounter-stainingwithVybrant

DyeCycleViolet.SignalswereanalysedusingaSMLMsetup.

a-b:untreatedcells;d-e:cellsafteronehourofONDstress.

b-e:magnificationsofwhiteboxesinaandd,respectively.

c-f:wide-fieldimagescorrespondingtobande,respectively.Regionsdeprivedof

chromatinarehighlightedwithanasteriskandatollsareshownwithawhitearrow.

Kirmesetal.GenomeBiology.2015.

染色质的构象和密度会随着细胞周期或环境的改变而改变。

染色质的组装密度能影响基因表达

2

透射电镜检测染色质构象实验的原理:

电镜检测染色质构象的操作流程:

33

取材:组织块约1mm,细胞沉淀体积约1mm。

固定:2.5%戊二醛,4℃固定过夜;0.1MPB(pH7.4),4℃漂洗10-15min×3次,

期间每5min颠倒几次;2%锇酸室温固定1.5h;0.1MPB(pH7.4)漂洗10-

15min×3次。

脱水:30%、50%、70%、80%、95%乙醇各10-15min;100%乙醇15min×3次;

100%丙酮15min×2次

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