遗传 ›› 1997, Vol. 19 ›› Issue (2): 1-4.

• 论文 •    下一篇

四甲基氯化铵在PCR扩增小麦基因中的关键作用

廖玉才1; 李和平1; Rainer Fischer2 LIAO Yu-Cai1;LI He-Ping1; Rainer Fischer2   

  1. 1.华中农业大学作物遗传改良国家重点实验室, 武汉 430070 2.德国亚琛技术大学植物及分子遗传研究所,亚琛52056 1.State Key Laboratory of Crop Genetic Improvement,Huazhaong Agricultural University,Wuhan 430070 2.Institute of Biology I(Botanik/Molekulargenetik),RWTH Aachen,D-52056 Aachen,Germany
  • 收稿日期:1900-01-01 出版日期:1997-04-10 发布日期:1997-04-10

Essential Role of Tetramethyl Ammonium Chloride for Amplification of Wheat Gene Sequences by Polymerase Chain Reaction (PCR)

  • Received:1900-01-01 Online:1997-04-10 Published:1997-04-10

摘要: 利用高简并性引物,用PCR法从小麦DNA或cDNA中合成小麦几丁质酶基因、葡
聚糖酶基因和苯丙氨酸解氨酶基因片段。在PCR反应中添加四甲基氯化铵(TMACl)是合成这些特异基因片段的关键。合成的PCR片段都经末端补齐和磷酸化后用于克隆。核酸序列分析证实,这些PCR产物分别与用于设计PCR引物的基因具有高度的同源性。
Abstract:In the presence of tetramethy1 ammonium chloride(TMAC1),a chitinase gene sequence,a phenylalanine ammonia-lyase gene sequence and a glucanase cDNA sequence of wheat were amplified with highly degenerate primers by PCR.The inclusion of TMAC1 in the PCR reactions was essential for successful amplification of the desired sequences from genomic DNA or cDNA in wheat.The ends of the PCR fragments were made flush and phosphorylated prior to cloning.Sequence analyses of the above PCR fragments confirmed their identities,showing high sequence similarities to the genes used for the design of PCR primers.

关键词: PCR, 四甲基氯化铵, 葡聚糖酶, 苯丙氨酸解氨酶

Key words,
几丁质酶, X染色体, 人类基因组, 物理图谱, YAC重叠群, STS图谱

Key words: Physical map, X chromosome, STS, Human genome, YAC contig