PCR-RFLP genotyping assay for a lactase persistence polymorphism upstream of the lactase-phlorizin hydrolase gene.
PCR-RFLP genotyping assay for a lactase persistence polymorphism upstream of the lactase-phlorizin hydrolase gene.
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PCR-RFLP 基因分型测定乳糖酶-根皮苷水解酶基因上游的乳糖酶持久性多态性。
DOI：
10.1089/gte.2004.8.190
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Sibley,Eric
中科院分区：
文献类型：
--
作者：
Chao,ChristinaK;Sibley,Eric
世界上大多数人在成熟过程中经历乳糖酶基因表达的下降，即所谓的乳糖酶不持久性。因此，乳糖酶不持久的成年人容易出现乳糖不耐症的症状。相比之下，乳糖酶持续性是一种常染色体显性遗传疾病，导致整个成年期乳糖酶基因表达水平高，乳糖耐受性持续。乳糖酶持久性最近与位于乳糖酶结构基因上游13，910个碱基的单核苷酸遗传变异（C → T突变）相关。我们的目的是开发一种限制性片段长度多态性（RFLP）的方法来检测C/T变异的一种手段，确定个人的遗传倾向于乳糖酶持久性或非持久性。用设计用于避免或突变相邻限制性位点的独特引物对213，910-bp变体位点周围的210-bp区域中的基因组DNA进行PCR扩增。用限制性内切酶CviJI消化扩增的DNA，该酶识别乳糖酶非持久性变体产生的碱基对序列。限制性酶切凝胶分析产生的DNA片段的预期诊断分子量大小的个体是纯合子或杂合子的乳糖酶持久性和nonpersistence的变种。通过DNA序列分析证实了基于RFLP的方法预测的基因型。基于RFLP的方法提供了一种快速和非侵入性的手段，分子检测的存在或不存在的乳糖酶持久性变异。
The majority of the world’s human population experiences a decline of lactase gene expression during maturation, so-called lactase nonpersistence. Thus, adults with lactase nonpersistence are susceptible to developing symptoms of lactose intolerance. By contrast, lactase persistence is an autosomal dominant heritable condition that results in a high level of lactase gene expression throughout adulthood and sustained lactose tolerance. Lactase persistence has recently been correlated with a single nucleotide genetic variant (a C → T mutation) located 13,910 bases upstream from the lactase structural gene. We aimed to develop a restriction fragment length polymorphism (RFLP) method of detecting the C/T variants as a means of identifying individuals genetically inclined toward lactase persistence or nonpersistence. Genomic DNA in a 210-bp region surrounding the 213,910-bp variant site was PCR amplified with unique primers designed to avoid or mutate adjacent restriction sites. The amplified DNA was digested with a restriction enzyme, CviJI, that recognizes the base pair sequence generated by the lactase nonpersistence variant. Restriction digest gel analysis yielded DNA fragments of the expected diagnostic molecular weight sizes for individuals that were homozygote or heterozygote for the lactase persistence and nonpersistence variants. The genotypes predicted by the RFLP-based method were confirmed by DNA sequence analysis. The RFLP-based method provides a quick and noninvasive means of molecular detection of the presence or absence of the lactase persistence variant.
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影响因子：
1.9
作者：
Büning, C;Ockenga, J;Schmidt, H
通讯作者：
Schmidt, H
乳糖酶持久性/非持久性多态性由顺式作用元件控制。
影响因子：
3.5
作者：
Y. Wang;C. Harvey;W. S. Pratt;V. Sams;M. Sarner;M. Rossi;S. Auricchio;D. Swallow
通讯作者：
D. Swallow
影响因子：
3.5
作者：
Harvey, CB;Wang, YX;Swallow, DM
通讯作者：
Swallow, DM
DOI：
10.1080/00365519850186607
发表时间：
1998-05-01
影响因子：
2.1
作者：
Peuhkuri, K;Poussa, T;Korpela, R
通讯作者：
Korpela, R
影响因子：
1.9
作者：
M.D. J. E. Losanoff;M. Richman;Myron D. Jones
通讯作者：
Myron D. Jones