Restriction isotyping of human apolipoprotein E by gene amplification and cleavage with HhaI.
Restriction isotyping of human apolipoprotein E by gene amplification and cleavage with HhaI.
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发表时间：
1990-03
影响因子：
6.5
通讯作者：
J. Hixson;D. T. Vernier
中科院分区：
文献类型：
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作者：
J. Hixson;D. T. Vernier
我们使用了限制性同种型分析（限制性内切酶同种型基因分型）对常见的载脂蛋白E同种型（E2、E3、E4）进行快速分型。载脂蛋白E限制性同种型分析使用寡核苷酸扩增包含氨基酸位置112和158的载脂蛋白E基因序列。扩增产物用HhaI酶切，并在聚丙烯酰胺凝胶上进行电泳。每种同种型都通过HhaI片段大小的独特组合来区分，这使得能够对所有纯合子和杂合子组合进行明确分型。HhaI在编码112精氨酸（E4）和158精氨酸（E3、E4）的GCGC处切割，但不在编码112半胱氨酸（E2、E3）和158半胱氨酸（E2）的GTGC处切割。
We have used restriction isotyping (restriction enzyme isoform genotyping) for rapid typing of common apolipoprotein E isoforms (E2, E3, E4). ApoE restriction isotyping used oligonucleotides to amplify apolipoprotein E gene sequences containing amino acid positions 112 and 158. The amplification products were digested with HhaI and subjected to electrophoresis on polyacrylamide gels. Each of the isoforms was distinguished by a unique combination of HhaI fragment sizes that enabled unambiguous typing of all homozygotic and heterozygotic combinations. HhaI cleaves at GCGC encoding 112arg (E4) and 158arg (E3, E4), but does not cut at GTGC encoding 112cys (E2, E3) and 158cys (E2).