Generating of rice OsCENH3-GFP transgenic plants and their genetic applications
Generating of rice OsCENH3-GFP transgenic plants and their genetic applications
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水稻OsCENH3-GFP转基因植株的产生及其遗传应用
DOI：
10.1007/s11434-008-0376-4
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发表时间：
2008-08
期刊：
影响因子：
--
通讯作者：
Cheng ZhuKuan
中科院分区：
文献类型：
--
作者：
Yu HengXiu;Gong ZhiYun;Wang Xin;Tang Ding;Gu MingHong;Cheng ZhuKuan
为了研究水稻功能着丝粒，构建了OsCENH 3-GFP嵌合基因，并通过农杆菌介导法将其转化到籼稻品种中籼3037中。通过PCR和Southern杂交证实外源基因已整合到转基因植株中。转基因植株与中籼3037一样，在整个生育期内生长正常。转基因植株的有丝分裂和减数分裂均未检测到明显缺陷。在T0和T1代植物的有丝分裂和减数分裂细胞中，GFP信号和抗CENH 3焦点的重叠表明GFP已经成功地与CENH 3融合，因此GFP信号可以很好地代表CENH 3在每条染色体上的位置。为了评价转基因植株在其他遗传学研究中的适用性，以水稻着丝粒串联重复序列CentO为探针，对花粉母细胞（PMCs）的偶线期染色体进行荧光原位杂交（FISH）。结果表明，GFP信号与CentO FISH信号重叠，表明CentO是构成水稻功能着丝粒的关键元件之一。抗α-tublin抗体和抗PAIR 2抗体在转基因植株有丝分裂和减数分裂期间的染色体上的免疫荧光染色进一步表明，OsCENH 3-GFP转基因植株可广泛用于水稻分子生物学研究，特别是用于活细胞和组织中功能着丝粒的标记。
In order to investigate rice functional centromeres, OsCENH3-GFP chimeric gene was constructed and transformed into the indica rice variety, Zhongxian 3037, mediated by Agrobacturium. The integration of the exogenous genes in the transgenic plants was confirmed by PCR and Southern blotting. The transgenic plants grow normally during their whole life time, just like Zhongxian 3037. No significant defects were detected in either mitosis or meiosis of the transgenic plants. The overlapping of GFP signals and anti-CENH3 foci in both mitotic and meiotic cells from T0 and T1 generation plants indicated that GFP had been successfully fused with CENH3, so the GFP signals can well represent the CENH3 locations on each chromosome. To evaluate the applicability of the transgenic plants to other genetic studies, fluorescence in situ hybridization (FISH) using rice centromeric tandem repetitive sequence CentO as the probe was conducted on the zygotene chromosomes of pollen mother cells (PMCs). It has been revealed that the GFP signals are overlapping with CentO FISH signals, showing that CentO is one of the key elements constituting rice functional centromeres. Immunofluorescent staining using anti-α-tublin antibody and anti-PAIR2 antibody on the chromosomes during mitosis and meiosis stages of the transgenic plants further reveals that OsCENH3-GFP transgenic plants can be widely used for studying rice molecular biology, especially for tagging functional centromeres in both living cells and tissues.
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影响因子：
3.3
作者：
Dongfen Zhang;Qiuying Yang;Weidong Bao;Yu Zhang;B. Han;Yongbiao Xue;Zhukuan Cheng
通讯作者：
Dongfen Zhang;Qiuying Yang;Weidong Bao;Yu Zhang;B. Han;Yongbiao Xue;Zhukuan Cheng
DOI：
10.1073/pnas.88.9.3734
发表时间：
1991-05-01
影响因子：
11.1
作者：
PALMER, DK;ODAY, K;MARGOLIS, RL
通讯作者：
MARGOLIS, RL
影响因子：
56.9
作者：
Schueler, MG;Higgins, AW;Willard, HF
通讯作者：
Willard, HF
影响因子：
11.6
作者：
Lermontova, Inna;Schubert, Veit;Schubert, Ingo
通讯作者：
Schubert, Ingo
影响因子：
4.1
作者：
Hosouchi, T;Kumekawa, N;Kotani, H
通讯作者：
Kotani, H