Amplification of a variable number of tandem repeats (VNTR) locus (pMCT118) by the polymerase chain reaction (PCR) and its application to forensic science.
Amplification of a variable number of tandem repeats (VNTR) locus (pMCT118) by the polymerase chain reaction (PCR) and its application to forensic science.
复制标题
通过聚合酶链式反应 (PCR) 扩增可变数量串联重复 (VNTR) 基因座 (pMCT118) 及其在法医学中的应用。
DOI：
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发表时间：
1990
影响因子：
1.6
通讯作者：
R. White
中科院分区：
文献类型：
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作者：
K. Kasai;Y. Nakamura;R. White
遗传基因座（由DNA探针PMCT118定义的D1S58）已通过聚合酶链（PCR）成功地从很少的基因组脱氧核酸（DNA）中成功扩增了包含可变数量的串联重复序列（VNTR）的遗传基因座。确定了基因座的DNA序列，并发现由16碱基共有序列和侧翼序列组成。合成了与侧翼序列互补的寡核苷酸引物作为通过PCR方法扩增MCT118的引物。从血液中分离出的人类基因组DNA（从每个样品中分离出2 ng）在MCT118基因座上成功扩增，并且在聚丙烯酰胺凝胶上电泳后可通过溴化乙锭染色检测多态性带。现在可以在24小时内常规实现该VNTR基因座的基因型的测定，而无需南部印迹或放射性材料。此外，在PCR扩增中产生的DNA片段的小尺寸（387至723个碱基对）允许单个等位基因仅通过一个重复单位而良好的单个等位基因。每个等位基因片段中存在的串联重复数量的精确规范可以从一个分析到另一个分析。
A genetic locus (D1S58, defined by DNA probe pMCT118) that contains a variable number of tandem repeats (VNTR) has been successfully amplified from a very small amount of genomic deoxyribonucleic acid (DNA) by the polymerase chain reaction (PCR). The DNA sequence of the locus was determined and was found to consist of a 16-base consensus sequence and flanking sequences. Oligonucleotide primers complementary to the flanking sequences were synthesized to serve as primers for amplification of MCT118 by the PCR method. Human genomic DNA isolated from blood (2 ng from each sample) was successfully amplified at the MCT118 locus, and polymorphic bands were detectable by ethidium bromide staining after electrophoresis on polyacrylamide gels. Determination of genotypes at this VNTR locus can now be routinely achieved within 24 h, without the need for Southern blots or radioactive materials. Furthermore, the small size (387 to 723 base pairs) of the DNA fragments produced in the PCR amplification permits good resolution of individual alleles that differ by only one repeat unit. The precise specification of the number of tandem repeats present in each allelic fragment is reproducible from one analysis to another.