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利用荧光原位杂交技术对六倍体栽培燕麦(Avena sativa L.)中共同转化的转基因进行染色体定位。

Chromosomal localization of cotransformed transgenes in the hexaploid cultivated oat Avena sativa L. using fluorescence in situ hybridization.

作者信息

Leggett J M, Perret S J, Harper J, Morris P

机构信息

Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, Ceredigion SY23 3EB, UK.

出版信息

Heredity (Edinb). 2000 Jan;84 ( Pt 1):46-53. doi: 10.1046/j.1365-2540.2000.00627.x.

Abstract

Fluorescence in situ hybridization (FISH) was used to localize two transgenes (gus and bar), carried on plasmids pACT-1F and pUBA, respectively, on mitotic metaphase squashes of T1 plants of the cultivated hexaploid oat Avena sativa L. cotransformed by microprojectile bombardment of embryogenic callus. Among the eight progeny analysed by FISH in each of two lines, we detected plants null, hetero- and homozygous for the two genes in one line, and plants null and heterozygous for the two genes in the other line. Our results demonstrated that in the two independent transformation events, the gus and bar genes had inserted in the same position relative to each other. In each transformation event, the insertions occurred on D satellite (SAT) chromosomes bearing a C genome translocation.

摘要

荧光原位杂交(FISH)用于将分别携带在质粒pACT - 1F和pUBA上的两个转基因(gus和bar)定位到通过胚性愈伤组织微粒轰击共转化的栽培六倍体燕麦 Avena sativa L. T1代植株的有丝分裂中期压片中。在对两个株系中每个株系的八个后代进行FISH分析时,我们在一个株系中检测到了两个基因的缺失、杂合和纯合植株,在另一个株系中检测到了两个基因的缺失和杂合植株。我们的结果表明,在这两个独立的转化事件中,gus和bar基因相对于彼此插入到了相同位置。在每个转化事件中,插入发生在带有C基因组易位的D卫星(SAT)染色体上。

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