Nishimura A, Tsuhako M, Miki T, Ogihara T, Baba Y
Department of Chemistry, Kobe Pharmaceutical University, Japan.
Chem Pharm Bull (Tokyo). 1998 Feb;46(2):294-7. doi: 10.1248/cpb.46.294.
Capillary electrophoresis with laser-induced fluorescence detector (CE-LIF) was successfully applied to the analysis of the variable number of tandem repeats (VNTR) in a human apolipoprotein B gene (APOB). Apolipoprotein B VNTR alleles containing more than 35 repeat units are a significant risk factor for heart disease. Thus, we developed a method for accurately determining the number of repeat units (16 bp) in the VNTR using capillary electrophoresis. The CE-LIF technique gave excellent resolution of APOB alleles differing by 2 or 4 repeat units over the range 600 to 1000 bp. The recommended conditions for the analysis of APOB VNTR loci by CE-LIF are as follows: effective length of capillary (100 microns i.d., 360 microns o.d.), 50 cm; running buffer, 50 mM Tris-borate 0.5% methylcellulose and 0.1 microM fluorescent dye YO-PRO-1; electric field, 150 V/cm.
采用带有激光诱导荧光检测器的毛细管电泳技术（CE-LIF）成功地对人类载脂蛋白B基因（APOB）中的可变串联重复序列（VNTR）进行了分析。含有超过35个重复单元的载脂蛋白B VNTR等位基因是心脏病的一个重要危险因素。因此，我们开发了一种利用毛细管电泳精确测定VNTR中重复单元数量（16 bp）的方法。CE-LIF技术在600至1000 bp范围内对相差2个或4个重复单元的APOB等位基因具有出色的分辨率。通过CE-LIF分析APOB VNTR位点的推荐条件如下：毛细管有效长度（内径100微米，外径360微米），50厘米；运行缓冲液，50 mM Tris-硼酸盐、0.5%甲基纤维素和0.1 microM荧光染料YO-PRO-1；电场，150 V/cm。
