辐射诱导染色体畸变的快速fish方法的建立
Keywords: 泛着丝粒探针,荧光原位杂交,染色体畸变,简并引物pcr
Abstract:
?目的:建立一种检测辐射诱导染色体畸变的快速荧光原位杂交(fish)方法。方法:两步简并引物pcr法扩增人α-卫星dna,制备荧光基团直接标记的泛着丝粒探针;对60coγ射线照射后的人外周血淋巴细胞染色体标本进行fish分析,荧光显微镜下检测着丝粒及染色体形态。结果:直接标记泛着丝粒探针杂交后显示所有染色体着丝粒均有较强的信号;应用制备的fish探针检测到γ射线照射诱导的双着丝粒、着丝粒环和易位染色体畸变,37℃杂交5-12h后经过简单的洗涤即可在荧光显微镜下检测到较好的信号。结论:本研究制备的直接标记泛着丝粒探针,可用于fish快速检测辐射诱导的染色体双着丝粒体、着丝粒环和易位畸变。
References
[1] | 刘青杰,陆雪,陈德清,等.《荧光原位杂交分析染色体易位估算辐射生物剂量技术方法》解读[j].国际放射医学核医学杂志,2012,36(4):245-248.
|
[2] | weierh-ug,lucasjn,poggenseem,etal.two-colorhybridizationwithhighcomplexitychromosome-specificprobesandadegeneratealphasatelliteprobednaallowsunambiguousdiscriminationbetweensymmetricalandasymmetricaltranslocations[j].chromosoma,1991,100(6):371-376.
|
[3] | chendq,zhangcy.asimpleandconvenientmethodforgainingpurepopulationsoflymphocytesatthefirstmitoticdivisioninvitro[j].mutatres,1992,282(3):227-229.
|
[4] | 卢圣栋.现代分子生物学实验技术[m].北京:高等教育出版社,1993:159.
|
[5] | 董玉玮,邱龙,李培青,等.pcr和随机引物标记探针的方法比较[j].生物学杂志,2007,24(1):63-66.
|
[6] | johnsonkl,nathj,pluthjm,etal.thedistributionofchromosomedamage,non-reciprocaltranslocationsandcolonialaberrationsinlymphocytesfromchemobylclear-upworkers[j].mutatres,1999,439(1):77-85.
|
[7] | haydenke,stromeed,merrettsl,etal.sequenceassociatedwithcentromerecometencyinhumangenome[j].molcellbiol,2013,33(4):763-772.
|
[8] | lamadridai,garciao,delbosm,etal.pcc-ringinductioninhumanlymphocytesexposedtogammaandneutronirradiation[j].jradiatres,2007,48(1):1-6.
|
[9] | 赵骅,陆雪,陈德清,等.早熟染色体凝聚最长染色体长宽比和最长与最短染色体长度比作为估算辐射剂量指标的研究[j].癌变·畸变·突变,2011,23(2):137-140.
|
[10] | mosessop,palittif,pepeg,etal.relationshipbetweenchromatinstructure,dnadamageandrepairfollowingx-irradiationofhumanlymphocytes[j].mutatres,2010,701(1):86-91.
|
[11] | willardhf.chromosome-specificorganizationofhumanalphasatellitedna[j].amjhumgenet,1985,37(3):524-532.
|
[12] | virala,fenechm,thierensh.themicronucleusassayasabiologicaldosimeterofinviroionizingradiationesposure[j].mutagenesis,2011,26(1):11-17.
|
[13] | baeyensa,swansonr,herdo,etal.asemi-automatedmicronucleus-centromereassaytoassesslow-doseradiationexposureinhumanlymphocytes[j].intjradiatbiol,2011,87(9):923-931.
|
[14] | baumgarthera,weierjf,weierhu.chromosome-specificdnarepeatprobes[j].jhistochemcytochem,2006,54(12):1363-1370.
|
Full-Text