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拟南芥AGL15基因的克隆及其植物超表达载体的构建 被引量:1
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作者 吴晓梅 刘忠丽 《河南农业科学》 CSCD 北大核心 2012年第5期33-36,共4页
为了研究AGL15基因在种子发育过程中的作用,应用聚合酶链式反应技术(PCR)扩增了拟南芥AGL15基因,并将其克隆到pMD20-T vector载体上,对重组子进行PCR检测和限制性内切酶分析,并测定了该基因全序列。结果表明,该基因全长为807bp。将拟南... 为了研究AGL15基因在种子发育过程中的作用,应用聚合酶链式反应技术(PCR)扩增了拟南芥AGL15基因,并将其克隆到pMD20-T vector载体上,对重组子进行PCR检测和限制性内切酶分析,并测定了该基因全序列。结果表明,该基因全长为807bp。将拟南芥AGL15基因定向克隆到植物表达载体pCambia1301的35S启动子下游,双酶切鉴定结果表明,AGL15基因在双元表达载体中的插入位置和方向都正确,即成功构建了该基因的植物超表达载体。 展开更多
关键词 拟南芥 PCR agl15基因 超表达载体
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基于TILLING技术筛选大豆GmAGL15基因突变体 被引量:1
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作者 孙彦波 李忠峰 +1 位作者 王俊 邱丽娟 《大豆科学》 CAS CSCD 北大核心 2019年第6期906-913,共8页
AGL15是MADS(MCM1-AGAMOUS-DEFICIENS-SRF)基因家族中的一员,MADS蛋白通过与下游基因启动子的顺式作用元件特异性结合调控植物种子中贮藏物质的积累。为验证大豆中AGL15基因的功能,利用TILLING技术在EMS诱变的中品661的M5中筛选出8个GmA... AGL15是MADS(MCM1-AGAMOUS-DEFICIENS-SRF)基因家族中的一员,MADS蛋白通过与下游基因启动子的顺式作用元件特异性结合调控植物种子中贮藏物质的积累。为验证大豆中AGL15基因的功能,利用TILLING技术在EMS诱变的中品661的M5中筛选出8个GmAGL15突变体。共检测到10个突变位点,其中6个突变位点发生在基因的编码区导致氨基酸发生改变,4个突变位点发生在基因的内含子。1个SNP导致该基因第2个外显子的第54个碱基由C变为T,并使该基因编码的第79位氨基酸由丙氨酸突变为缬氨酸;5个SNP发生在该基因的第6个外显子的112位碱基上,使得碱基由T变为A,导致该基因编码的第568位氨基酸由亮氨酸突变为甲硫氨酸。大豆GmAGL15的突变分布频率为1/805 kb。考种发现5个非同义突变体和1个内含子突变体的蛋白质或脂肪含量与野生型ZP661相比差异显著。本研究发现的等位变异有助于阐明GmAGL15在大豆种子中的作用机制,并为大豆的遗传改良提供宝贵的种质资源。 展开更多
关键词 大豆 转录因子 agl15 EMS TILLING
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Characterization of Arabidopsis MYB transcription factor gene AtMYB17 and its possible regulation by LEAFY and AGL15 被引量:24
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作者 Yunfei Zhang Guangyu Cao Li-Jia Qu Hongya Gu 《Journal of Genetics and Genomics》 SCIE CAS CSCD 2009年第2期99-107,共9页
MYB transcription factors compose one of the largest transcription factor families in Arabidopsis, which play important roles in vari- ous developmental processes as well as defense responses against environmental str... MYB transcription factors compose one of the largest transcription factor families in Arabidopsis, which play important roles in vari- ous developmental processes as well as defense responses against environmental stresses. In this study, we report the characterization of AtMYB17 gene, a putative R2R3 type MYB gene family member in Arabidopsis. AtMYB17 was found exclusively localized in nuclear, with an activation domain at its C-terminus. AtMYBI7 was highly expressed in inflorescences and siliques, especially at early flower developmental stages. The level of AtMYB17 transcripts was also found to increase after imbibition during seed germination and gradually concentrate to the shoot apex. Bioinformatics analysis identified several binding sites of LEAFY (LFY) and AGL15 in the promoter re- gion of AtMYB17. Promoter-GUS fusion analysis showed that the LFY binding sites were important in fine-tuning regulation of the spatio-temporal expression ofAtMYB17 in transgenic plants. Moreover, AtMYB17 was up-regulated in 35S::AGL15 plants. Taken together, our data suggest that LFY may be involved in the regulation of AtMYB17, possibly together with AGL 15, and thereafter in early inflores- cence development and seed germination. 展开更多
关键词 AtMYB17 transcription factor ARABIDOPSIS LEAFY agl 15
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灰毡毛忍冬内参基因筛选和Mads-box家族基因AGL15的时空表达分析 被引量:10
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作者 蔡嘉洛 朱贻霖 +7 位作者 谢舒平 陈林 欧阳琳 刘峰 张亚丽 刘湘丹 童巧珍 易刚强 《中草药》 CAS CSCD 北大核心 2016年第15期2727-2733,共7页
目的筛选灰毡毛忍冬Lonicera macranthoides基因表达分析的内参基因,研究其Mads-box基因家族AGL15基因(Lm AGL15)的时空表达特性。方法克隆灰毡毛忍冬内参基因18 S r RNA、Ubiquilin、Actin和Efl-β的基因片段,评价了4个基因在不同部位... 目的筛选灰毡毛忍冬Lonicera macranthoides基因表达分析的内参基因,研究其Mads-box基因家族AGL15基因(Lm AGL15)的时空表达特性。方法克隆灰毡毛忍冬内参基因18 S r RNA、Ubiquilin、Actin和Efl-β的基因片段,评价了4个基因在不同部位叶、茎和花蕾,以及4个基因在灰毡毛忍冬生长不同时期的稳定性,筛选内参基因,分析Lm AGL15基因的时空表达特性。结果 18 S r RNA表达最稳定,适宜作为内参基因,叶片、茎的Lm AGL15基因相对表达量较低,花蕾相对表达量较高。花蕾不同发育时期,Lm AGL15的相对表达量呈现逐渐降低的变化,20 d后Lm AGL15的相对表达量最低,然后依次是花蕾初期(15 d)、青绿色花蕾期(10 d)、绿白色花蕾期(5 d)。结论 18 S r RNA是适宜的内参基因。叶片、茎和花蕾中,Lm AGL15的相对表达量差异明显。花蕾发育不同时期,Lm AGL15的相对表达量呈逐渐降低的变化,与花蕾开放变化规律相似。 展开更多
关键词 灰毡毛忍冬 内参基因 蕾期延长 时空表达特性 18 S r RNA UBIQUILIN ACTIN Efl-β agl15
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Two MADS-box proteins,AGL9 and AGL15,recruit the FIS-PRC2 complex to trigger the phase transition from endosperm proliferation to embryo development in Arabidopsis
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作者 Shen Zhang Devasantosh Mohanty +1 位作者 Adnan Muzaffar Min Ni 《Molecular Plant》 SCIE 2024年第7期1110-1128,共19页
Spatiotemporal regulation of gene expression by polycomb repressive complex 2(PRc2)is critical for animal and plant development.The Arabidopsis fertilization independent seed(FIS)-PRC2 complex functions specifically d... Spatiotemporal regulation of gene expression by polycomb repressive complex 2(PRc2)is critical for animal and plant development.The Arabidopsis fertilization independent seed(FIS)-PRC2 complex functions specifically during plant reproduction from gametogenesis to seed development.After a double fertilization event,triploid endosperm proliferates early,followed by the growth of a diploid embryo,which replaces the endosperm in Arabidopsis and many dicots.Key genes critical for endosperm proliferation such as IKkU2 and MINI3 are activated after fertilization.Here we report that two MADS-box AGAMOUS-LIKE(AGL)proteins associate with the key endosperm proliferation loci and recruit the FIS-PRC2 repressive complex at 4-5 days after pollination(DAP).Interestingly,AGL9 and AGL15 only accumulate toward the end of endosperm proliferation at 4-5 DAP and promote the deposition of H3K27me3 marks at key endosperm proliferation loci.Disruption of AGL9 and AGL15 or overexpression of AGL9 or AGL15 significantly influence endosperm proliferation and cellularization.Genome-wide analysis with cleavage Under Targets and tagmentation(CUT&Tag)sequencing and RNA sequencing revealed the landscape of endosperm H3K27me3 marks and gene expression profiles in Col-0 and agl9 agl15.CUT&Tag qPCR also demonstrated the occupancy of the two MADS-box proteins and FIS-PRC2 on a few representative target loci.Our studies suggest that MADS-box proteins could potentially recruit PRc2 to regulate many other developmental processes in plants or even in fungi and animals. 展开更多
关键词 agl9 agl15 embryo endosperm IKU2 MEDEA MINI3
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