Searching for gene defects by denaturing gradient gel electrophoresis.
Searching for gene defects by denaturing gradient gel electrophoresis.
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通过变性梯度凝胶电泳寻找基因缺陷。
DOI：
10.1101/sqb.1986.051.01.034
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Grinfeld,E
中科院分区：
文献类型：
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作者：
Lerman,LS;Silverstein,K;Grinfeld,E
在试图确定人类胎儿中关键基因的特定等位基因的存在时，最希望知道其基因组的精确碱基序列。目前尚不清楚直接测序技术何时能变得足够快速和简单，以允许在诊断相关分组（DRGs）设定的医疗保险报销范围内审查50 kb，但目前，其他检测偏离正常序列的途径似乎是必要的。显性等位基因或性连锁等位基因的存在可以通过与适当序列标记的连锁在一个家族中进行追踪。目前的实践主要依赖于限制性片段长度多态性（RFLPs）作为标记，但简单的高可变重复序列也很有希望。胎儿受到影响的可能性的估计取决于有多少其他家庭成员可以接受检测，以及基因和标记之间联系的密切程度。对于基因产物没有生化检测的隐性基因，必须通过两个家族来追踪与附近标记的联系。如果携带有性联系缺陷的雄性不太可能繁殖，连锁分析的帮助有限；三分之一的缺陷代表新的突变。只需要测试一个人的DNA，而不是一个家庭的DNA，将从根本上改变测试的前景和得出可靠结论的可能性。
In attempting to ascertain the presence of particular alleles of critical genes in a human fetus, it would be most desirable to know precisely the base sequence of its genome. It is not clear how soon direct sequencing techniques will become fast and easy enough to permit scrutiny of 50 kb within the limits of Medicare reimbursement set by the diagnostic related groupings (DRGs), but for the present, other paths to detection of departures from normal sequence seem to be necessary.The presence of a dominant or sex-linked allele can be traced through a family by means of linkage to an appropriate sequence marker. Current practice depends mostly on restriction-fragment-length polymorphisms (RFLPs) as markers, but simple hypervariable repeats are also promising. The estimate of the probability that the fetus is affected depends on which and how many other family members are available for testing and on the closeness of the linkage between the gene and marker. For recessive genes where there is no biochemical test for a gene product, linkage to nearby markers must be traced through two families. Linkage analysis is of limited help where a male carrying a sexlinked defect is unlikely to reproduce; one third of all defects represent new mutations. The need to test the DNA of only one individual, rather than a family, would radically alter the outlook for testing and for a secure conclusion.