Detecting sequence changes in a gene.
Detecting sequence changes in a gene.
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检测基因中的序列变化。
DOI：
10.1007/bf01534942
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
Lerman,LS
中科院分区：
文献类型：
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作者：
Lerman,LS
只有从单个个体或胎儿细胞的DNA样本中推断出任何基因型的相关方面，我们对人类遗传学的理解才会带来最大的好处。虽然通过与多态标记的联系来追踪不受欢迎的等位基因作为当前实践中的权宜之计有时是决定性的，但更多的时候，它只提供了一个广泛的概率估计，这将是一个艰难的个人决策的机会指导。直接表征决定疾病或风险的基因是遗传研究的主要目标。根据目前的分子生物学标准，唯一完全令人满意的基因特征需要确定其碱基序列，如果它是常染色体，则需要确定两个代表。获得充分信息所需的序列长度可能包括长侧翼部分，而且可以想象碱基修饰的鉴定可能是有用的。显然，在目前的技术条件下，这种水平的分析是不可能作为常规程序的。A型血友病患者中有三分之一的患者携带新的突变基因，对其进行检测可能需要对105个碱基对进行测序，这可能需要在分离出携带该基因片段的克隆后一年的工作。在目前的手段下，在没有先验知识的情况下搜索序列变化的实用性充其量是边缘的。通过将寡核苷酸探针与Southern印迹结合的方法对血友病A进行全面检查可能需要进行与大约10种不同寡核苷酸结合的测试
The greatest benefits of our understanding of human genetics will not be realized until relevant aspects of any genotype can be inferred from a sample of DNA from the cells of a single individual or a fetus. While tracking undesirable alleles by means of their linkage to polymorphic markers as the expedient in current practice is sometimes decisive, more often it provides only a broad probability estimate that will be a chancy guide to difficult personal decisions. Direct characterization of genes determining illness or risk represents a major goal of genetic investigation. By present molecular biological criteria, the only fully satisfactory characterization of a gene requires determination of its base sequence and of both representatives if it is autosomal. The length of sequence necessary for adequate information might include long flanking sections, and it is also conceivable that identification of base modifications might be useful. Clearly, analysis at this level is out of the question as a routine procedure with current technology. A test for hemophilia A, where one third of the afflicted carry new mutations, could require sequencing of 10 5 base pairs, perhaps a year's work after clones carrying the fragments of the gene have been isolated.Within present means, the practicability of searching for a sequence alteration without a priori knowledge of its site is at best marginal. Complete scrutiny for hemophilia A by means of binding of oligonucleotide probes to Southern blots might require tests of binding with about 10 a different oligonucleotides, and