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麦类作物体细胞基因组原位杂交(GISH)效果影响因素的分析
引用本文:林小虎,李兴锋,王黎明,陆文辉,王洪刚.麦类作物体细胞基因组原位杂交(GISH)效果影响因素的分析[J].实验生物学报,2005,38(2):126-132.
作者姓名:林小虎  李兴锋  王黎明  陆文辉  王洪刚
作者单位:[1]山东农业大学农学院,国家小麦改良中心泰安分中心,泰安271018 [2]山东农业大学农学院,国家小麦改良中心泰安分中心,泰安271018//河北科技师范学院,秦皇岛066600 [3]山东农业大学农学院,国家小麦改良中心泰安分中心,泰安271018//河南科技大学,洛阳471003
摘    要:以八倍体小滨麦、八倍体小黑麦、八倍体小偃麦、小麦一中间偃麦草双体异附加系为实验材料对影响麦类作物体细胞GISH技术实验效果的因素进行分析,研究结果表明:细胞分裂相多、染色体分散良好、无杂质影响的高质量的染色体制片是取得理想实验效果的基础;探针DNA浓度与封阻DNA浓度的比例及杂交后洗脱条件的控制是取得理想实验效果的关键。此外,还对麦类作物体细胞基因组原位杂交实验中出现的染色体丢失、外源染色体无杂交信号、杂交信号的强弱、杂交信号过多(杂交背景重)或过少、噪音信号及杂交污点产生的原因进行了分析,并提出了相应的解决方法或注意事项。

关 键 词:麦类作物  基因组原位杂交  体细胞  DNA浓度  八倍体小滨麦  八倍体小黑麦  八倍体小偃麦  双体异附加系  杂交信号  理想实验  中间偃麦草  染色体制片  染色体丢失  外源染色体  GISH  实验材料  细胞分裂  研究结果  杂质影响  洗脱条件

Analysis of factors influencing GISH used in Triticeae crops' body cells]
Hiao Hu Lin,Xing Feng Li,Li Ming Wang,Wen Hui lu,Hong Gang Wang.Analysis of factors influencing GISH used in Triticeae crops'' body cells][J].Acta Biologiae Experimentalis Sinica,2005,38(2):126-132.
Authors:Hiao Hu Lin  Xing Feng Li  Li Ming Wang  Wen Hui lu  Hong Gang Wang
Institution:Agronomy College of Shandong Agricultural University, Subcentre of National Wheat Improvement Center, Taian, 271018.
Abstract:Factors influencing genomic in situ hybridization (GISH) used in the body cells of Triticeae crops including Octoploid Tritileymus, Octoploid Trititrigia, Octoploid Tritelytrigia and wheat-Elytrigia intermedium alien disomic addition line were analyzed. The results indicated that ideal GISH effects were based on the high quality chromosome preparations containing more cell mitotic metaphase, better dispersed chromosomes and no foreign matter. The critical factors were suitable ratio of probe DNA to blocker DNA, and the control of elution condition. In addition, some abnormal phenomena, such as chromosome loss, no hybridization signal in alien chromosome, strong or weak hybridization signal, poor or overabundant hybridization signal (high hybridization background), noisy hybridization signal and hybridization spots, were preliminarily analyzed. Proposals and solutions for these problems were further brought forth.
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