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四甲基氯化铵在PCR扩增小麦基因中的关键作用
引用本文:廖玉才,李和平,Rainer FischerLIAO Yu-Cai,LI He-Ping,Rainer Fischer.四甲基氯化铵在PCR扩增小麦基因中的关键作用[J].遗传,1997,19(2):1-4.
作者姓名:廖玉才  李和平  Rainer FischerLIAO Yu-Cai  LI He-Ping  Rainer Fischer
作者单位: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
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  

Essential Role of Tetramethyl Ammonium Chloride for Amplification of Wheat Gene Sequences by Polymerase Chain Reaction(PCR)
LIAO Yu-Cai,LI He-Ping,Rainer Fischer.Essential Role of Tetramethyl Ammonium Chloride for Amplification of Wheat Gene Sequences by Polymerase Chain Reaction(PCR)[J].Hereditas,1997,19(2):1-4.
Authors:LIAO Yu-Cai  LI He-Ping  Rainer Fischer
Abstract:In the presence of tetramethyl ammonium chloride (TMACl), 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 TMACl 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.
Keywords:PCR  TMACl  Chitinase  Glucanase  Phenylalanine ammonia-lyase  
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