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化学学报
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化学学报 ›› 2009, Vol. 67 ›› Issue (4): 323-328. 上一篇    下一篇

研究论文

毛细管电泳-限制性片段长度多态性分析检测胃癌H-ras基因点突变

刘圆圆a,b,c 王 荣*,a 贾正平a 郭志强a 辛晓婷a,c
谢 华a 马 骏a 王 娟a 李文斌a

  

  1. (a兰州军区兰州总医院临床药理基地 兰州 730050)
    (b甘肃省红十字血液中心 兰州 730046)
    (c兰州大学生命科学学院 兰州 730000)

  • 投稿日期:2008-01-02 修回日期:2008-08-27 发布日期:2009-02-28
  • 通讯作者: 王荣

H-ras Gene Mutation Detection of Gastric Cancer by Restriction Fragment Length Polymorphism with Capillary Electrophoresis

Liu, Yuanyuan a,b,c Wang, Rong *,a Jis, Zhengping a Guo, Zhiqiang a
Xin, Xiaoting a,c Xie, Hua a Ma, Jun a Wang, Juan a Li, Wenbin a

  

  1. (a Department of Pharmacy, Lanzhou General Hospital of PLA, Lanzhou 730050)
    (b Gansu Red Cross Blood Center, Lanzhou 730046)
    (c Life Science College, Lanzhou University, Lanzhou 730000)
  • Received:2008-01-02 Revised:2008-08-27 Published:2009-02-28
  • Contact: WANG Rong

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1201

被引次数

8

建立一种毛细管电泳快速高效检测限制性内切酶酶切产物的方法, 使其更好地用于基因诊断. 以甲基纤维素(Methyl cellulose, MC)为筛分介质, 用pUC19 DNA/Msp I (Hpa II) Marker标准DNA片段为实验对象, 通过考察筛分介质的浓度、pH值、毛细管的温度和运行电压优化出分离小于600 bp的双链DNA片段的最适条件, 并将此方法应用于临床59例胃癌患者肿瘤组织H-ras基因12位密码子点突变情况的检测. MC是一种良好的筛分介质, 运用其进行毛细管电泳对于遗传性疾病的诊断将更加快速、准确、简便、灵敏.

关键词: 毛细管电泳, 甲基纤维素, 限制性片段长度多态性, H-ras, 基因突变

To establish the high performance detection method for separating products of restrictive enzyme DNA fragments and diagnosing mutation gene of gastric cancer by restriction fragment length polymorphism with capillary electrophoresis, methyl cellulose was used to separate pUC19 DNA/Msp I (Hpa II) fragments through capillary electrophoresis as sieving matrix. The effects of the concentration of methyl cellulose solution, running pH, running temperature and electric field strength on dsDNA (＜600 bp) analysis were investigated in the experiment. The optimum separation condition was obtained and applied to mutation detection of H-ras gene at codon 12 of 59 gastric cancer tissues. Methyl cellulose is an excellent medium and it will make diagnosis of genetic disease rapidly, exactly, conveniently and sensitively.

Key words: capillary electrophoresis, methyl cellulose, restriction fragment length polymorphism, H-ras, gene mutation