Donor splice site mutation in the apolipoprotein (Apo) C-II gene (Apo C-IIHamburg) of a patient with Apo C-II deficiency.
Donor splice site mutation in the apolipoprotein (Apo) C-II gene (Apo C-IIHamburg) of a patient with Apo C-II deficiency.
复制标题
Apo C-II 缺陷患者的载脂蛋白 (Apo) C-II 基因 (Apo C-IIHamburg) 中的供体剪接位点突变。
DOI：
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发表时间：
1988
影响因子：
15.9
通讯作者：
H. Brewer
中科院分区：
文献类型：
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作者：
S. Fojo;U. Beisiegel;U. Beil;K. Higuchi;M. Bojanovski;R. Gregg;H. Greten;H. Brewer
在1例载脂蛋白C-Ⅱ缺乏症患者（载脂蛋白C-Ⅱ汉堡）中分析了载脂蛋白C-Ⅱ的DNA、RNA和蛋白质。通过免疫印迹和免疫组化分析证明血浆和肝内C-II载脂蛋白水平显著降低。北方、狭缝印迹和原位杂交研究显示正常大小的载脂蛋白C-II mRNA水平较低。Southern杂交未检测到载脂蛋白C-Ⅱ基因的主要重排。载脂蛋白C-II基因组克隆的序列分析显示，内含子II的供体剪接位点内存在G至C取代。这种碱基取代导致形成新的Dde I和Hph I限制性酶切位点的丧失。通过聚合酶链反应扩增突变序列，并用Dde I和Hph I限制性内切酶消化，确定该患者为G至C突变纯合子。本文首次报道了一例载脂蛋白C-Ⅱ缺乏症患者的载脂蛋白C-Ⅱ异常基因的DNA序列。我们认为，这种供体剪接位点突变是导致剪接缺陷的主要遗传缺陷，并最终导致该家系中载脂蛋白C-II缺乏。
The DNA, RNA, and protein of apo C-II have been analyzed in a patient with apo C-II deficiency (apo C-IIHamburg). Markedly reduced levels of plasma and intrahepatic C-II apolipoprotein were demonstrated by immunoblotting and immunohistochemical analysis. Northern, slot blot, and in situ hybridization studies revealed low levels of a normal-sized apo C-II mRNA. No major rearrangement of the apo C-II gene was detected by Southern blotting. Sequence analysis of apo C-II genomic clones revealed a G-to-C substitution within the donor splice site of intron II. This base substitution resulted in the formation of a new Dde I and loss of a Hph I restriction enzyme cleavage site. Amplification of the mutant sequence by the polymerase chain reaction and digestion with Dde I and Hph I restriction enzymes established that the patient was homozygous for the G-to-C mutation. This is the initial report of the DNA sequence of an abnormal apo C-II gene from a patient with deficiency of apo C-II. We propose that this donor splice site mutation is the primary genetic defect that leads to defective splicing and ultimately to an apo C-II deficiency in this kindred.