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陈春丽, 郭文武, 邓秀新. 柑橘基因组原位杂交(GISH)技术体系的建立[J]. 植物科学学报, 2003, 21(5): 439-443.
引用本文: 陈春丽, 郭文武, 邓秀新. 柑橘基因组原位杂交(GISH)技术体系的建立[J]. 植物科学学报, 2003, 21(5): 439-443.
CHEN Chun-Li, GUO Wen-Wu, DENG Xiu-Xin. Establishment of GISH Technique in Citrus[J]. Plant Science Journal, 2003, 21(5): 439-443.
Citation: CHEN Chun-Li, GUO Wen-Wu, DENG Xiu-Xin. Establishment of GISH Technique in Citrus[J]. Plant Science Journal, 2003, 21(5): 439-443.

柑橘基因组原位杂交(GISH)技术体系的建立

Establishment of GISH Technique in Citrus

  • 摘要: 通过改进实验方法,建立起柑橘基因组原位杂交(Genomic in situ hybridization,GISH)分析技术,并成功地应用于属间有性杂种鉴定,为进一步分析柑橘体细胞杂种核基因组组成奠定了基础。柑橘的染色体制片以去壁低渗-火焰干燥法较好,容易获得大量清晰、分散的有丝分裂中期相,且杂质较少。切刻平移法生物素(Biotin)标记探针,标记基因组总DNA与封阻基因组总DNA的浓度比例为1:50时,能有效分开属间杂种枳橙中来自双亲的染色体。

     

    Abstract: Genomic in situ hybridization (GISH) technique was established in Citrus to identify intergeneric sexual hybrid. It showed that the modified procedures could be further used to identify Citrus somatic hybrids. Flame-drying protocol after enzyme digestion of cell wall was a good method for getting clear and well dispersed Citrus chromosome preparation with few debris. With probe DNA labeled by nick translation, and the blocking DNA being 50 times the amount of the probe DNA, Carrizo citrange,an intergeneric sexual hybrid of Citrus, was successfully identified by GISH method.

     

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