ANALYSIS OF REPLICATION TIMING PROPERTIES OF HUMAN X-CHROMOSOMAL LOCI BY FLUORESCENCE IN-SITU HYBRIDIZATION
ANALYSIS OF REPLICATION TIMING PROPERTIES OF HUMAN X-CHROMOSOMAL LOCI BY FLUORESCENCE IN-SITU HYBRIDIZATION
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DOI：
10.1073/pnas.91.13.6083
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发表时间：
1994-06-21
影响因子：
11.1
通讯作者：
CHINAULT, C
中科院分区：
文献类型：
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作者：
BOGGS, BA;CHINAULT, C
我们在正常雌性细胞的间期细胞核上使用荧光原位杂交来比较人类X染色体上已知逃避X失活的基因和那些失活的基因的复制时间模式。通过这种方法，不仅可以确定不同基因座的较早复制等位基因的相对复制时间，还可以估计每个基因座的两个等位基因复制的异步程度。通常失活的基因座，如HPRT和FRAXA，显示出高度的复制异步，而未失活的基因座（ZFX和RPS4X）被发现复制非常同步。有趣的是，用这种方法检测仅从失活X染色体上表达的XIST，发现它也是异步复制的，表达的拷贝显然先复制。因此，通过检查X染色体上的不同位点，可以确定单个基因的表达与相对复制时间之间存在严格的相关性。
We have used fluorescence in situ hybridization on interphase nuclei of normal female cells to compare the replication timing patterns of genes on the human X chromosome that are known to escape X inactivation with those that are inactivated. By this procedure it was possible not only to determine the relative time of replication of the earlier-replicating allele for different loci but also to estimate the degree of asynchrony of replication of the two alleles for each individual locus. Loci such as HPRT and FRAXA, which are normally inactivated, displayed a high degree of replication asynchrony, whereas loci that are not inactivated (ZFX and RPS4X) were found to replicate very synchronously. Interestingly, examination of XIST, which if expressed only from the inactive X chromosome, by this procedure revealed that it also replicated asynchronously, with the expressed copy apparently replicating first. Therefore, by examining different loci from the X chromosome it was determined that there is a strict correlation between the expression and relative time of replication of individual genes.