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棉花中一个类DREB1/CBF基因(GhDREB1L)的分子克隆及其功能分析
黄波,金龙国,刘进元
中国科学 生命科学 , 2006,
Abstract: DREB1/CBF类转录因子在植物抵抗外界环境胁迫上起到非常重要的作用,而且对于利用基因工程技术来提高植物的抗逆性也非常有用.从棉花中分离出一个编码DREB1/CBF类蛋白(GhDREB1L)的cDNA,并对该蛋白质的序列特性、DNA结合特性及转录本的表达特征进行了分析.GhDREB1L含有一个保守的AP2/ERF结构域,其氨基酸序列与其他植物中DREB家族的DREB1/CBF组成员高度相似.采用Ni-NTA亲和层析技术,成功纯化了在BL21(DE3)菌株中表达的GhDREB1L蛋白的DNA结合区.凝胶滞留实验揭示,纯化的GhDREB1L融合蛋白能够特异性地与已经得到鉴定的DRE元件(核心区,ACCGAC)以及胚胎发育晚期丰富蛋白基因LEAD113启动子区中的类DRE元件(核心区,GCCGAC)结合.半定量RT-PCR显示GhDREB1L基因在棉花子叶中受到低温、干旱以及NaCl处理的诱导.这些结果暗示了棉花GhDREB1L转录因子可能是通过与DRE元件的结合,在低温、干旱及高盐的应答途径中起重要作用.
棉花中一个类DREB1/CBF基因(GhDREB1L)的分子克隆及其功能分析  [PDF]
黄波,金龙国,刘进元
中国科学 生命科学 , 2006,
Abstract: DREB1/CBF类转录因子在植物抵抗外界环境胁迫上起到非常重要的作用,而且对于利用基因工程技术来提高植物的抗逆性也非常有用.从棉花中分离出一个编码DREB1/CBF类蛋白(GhDREB1L)的cDNA,并对该蛋白质的序列特性、DNA结合特性及转录本的表达特征进行了分析.GhDREB1L含有一个保守的AP2/ERF结构域,其氨基酸序列与其他植物中DREB家族的DREB1/CBF组成员高度相似.采用Ni-NTA亲和层析技术,成功纯化了在BL21(DE3)菌株中表达的GhDREB1L蛋白的DNA结合区.凝胶滞留实验揭示,纯化的GhDREB1L融合蛋白能够特异性地与已经得到鉴定的DRE元件(核心区,ACCGAC)以及胚胎发育晚期丰富蛋白基因LEAD113启动子区中的类DRE元件(核心区,GCCGAC)结合.半定量RT-PCR显示GhDREB1L基因在棉花子叶中受到低温、干旱以及NaCl处理的诱导.这些结果暗示了棉花GhDREB1L转录因子可能是通过与DRE元件的结合,在低温、干旱及高盐的应答途径中起重要作用.
A C-Repeat Binding Factor Transcriptional Activator (CBF/DREB1) from European Bilberry (Vaccinium myrtillus) Induces Freezing Tolerance When Expressed in Arabidopsis thaliana  [PDF]
Rachael J. Oakenfull, Robert Baxter, Marc R. Knight
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0054119
Abstract: Freezing stress affects all plants from temperate zones to the poles. Global climate change means such freezing events are becoming less predictable. This in turn reduces the ability of plants to predict the approaching low temperatures and cold acclimate. This has consequences for crop yields and distribution of wild plant species. C-repeat binding factors (CBFs) are transcription factors previously shown to play a vital role in the acclimation process of Arabidopsis thaliana, controlling the expression of hundreds of genes whose products are necessary for freezing tolerance. Work in other plant species cements CBFs as key determinants in the trait of freezing tolerance in higher plants. To test the function of CBFs from highly freezing tolerant plants species we cloned and sequenced CBF transcription factors from three Vaccinium species (Vaccinium myrtillus, Vaccinium uliginosum and Vaccinium vitis-idaea) which we collected in the Arctic. We tested the activity of CBF transcription factors from the three Vaccinium species by producing transgenic Arabidopsis lines overexpressing them. Only the Vaccinium myrtillus CBF was able to substantially activate COR (CBF-target) gene expression in the absence of cold. Correspondingly, only the lines expressing the Vaccinium myrtillus CBF were constitutively freezing tolerant. The basis for the differences in potency of the three Vaccinium CBFs was tested by observing cellular localisation and protein levels. All three CBFs were correctly targeted to the nucleus, but Vaccinium uliginosum CBF appeared to be relatively unstable. The reasons for lack of potency for Vaccinium vitis-idaea CBF were not due to stability or targeting, and we speculate that this was due to altered transcription factor function.
Gene cloning and molecular breeding to improve fiber qualities in cotton
Wangzhen Guo,Jing Sun,Tianzhen Zhang
Chinese Science Bulletin , 2003, DOI: 10.1360/02wc0463
Abstract: Cotton fiber is one of known natural resources comprising the highest purity cellulose. It plays an important role worldwide in the textile industry. With the acceleration of spinning speeds and the improvement of the people’s living level, the demand of improving cotton fiber qualities is getting stronger and stronger. So, making clear the developmental model of fiber cell and elucidating systematically the molecular mechanisms of cotton fiber development and regulation will produce a great significance to make full use of cotton gene resources, raise cotton yield and improve fiber quality, and even develop man-made fiber. In the paper, the status of the gene cloning and the molecular breeding to improve cotton fiber quality were reviewed, the importance and potential of gene cloning related with cotton fiber quality were put forward and the proposal and prospect on fiber quality improvement were made. Using national resources available and through the creative exploration in corresponding research, some international leading patents in genes or markers linked with cotton fiber development having Chinese own intellectual property should be licensed quickly. And they can be used to improve cotton fiber quality in cotton breeding practice.
Gene cloning and molecular breeding to improve fiber qualities in cotton
GUO Wangzhen,SUN Jing,ZHANG Tianzhen,
GUOWangzhen
,SUNJing,ZHANGTianzhen

科学通报(英文版) , 2003,
Abstract: Cotton fiber is one of known natural resources comprising the highest purity cellulose. It plays an important role worldwide in the textile industry. With the acceleration of spinning speeds and the improvement of the peoples liv-ing level, the demand of improving cotton fiber qualities is getting stronger and stronger. So, making clear the develop-mental model of fiber cell and elucidating systematically the molecular mechanisms of cotton fiber development and regulation will produce a great significance to make full use of cotton gene resources, raise cotton yield and improve fiber quality, and even develop man-made fiber. In the paper, the status of the gene cloning and the molecular breeding to im-prove cotton fiber quality were reviewed, the importance and potential of gene cloning related with cotton fiber quality were put forward and the proposal and prospect on fiber quality improvement were made. Using national resources available and through the creative exploration in corre-sponding research, some international leading patents in genes or markers linked with cotton fiber development hav-ing Chinese own intellectual property should be licensed quickly. And they can be used to improve cotton fiber quality in cotton breeding practice.
Molecular cloning and characterization of a cotton phosphoenolpyruvate carboxylase gene

Zhixin Qiao,Jinyuan Liu,

自然科学进展 , 2008,
Abstract: Phosphoenolpyruvate carboxylase (PEPC) plays diverse physiological functions during plant development. In this study, a new phosphoenolpyruvate carboxylase gene GhPEPC2 is isolated from cotton (Gossypium hirsutum cv. zhongmian 35) by RACE-PCR. The cloned cDNA of GhPEPC2 is 3364 bp in length, and has an open reading frame of 2913 bp, encoding for 971 putative amino acids with a calculated molecular mass of 110.6 kD and pI of 5.56. The deduced amino acid sequence of GhPEPC2 shares high similarity with other reported plant PEPCs. Southern blot analysis indicates that the cotton PEPC exists as a small gene family and the GhPEPC2 might have two copies in the cotton genome. The semi-quantitative RT-PCR reveals that GhPEPC2 constitutively expresses in all the tissues of cotton and accumulated highly in roots, flowers and embryos but relatively low in stems and fibers. In addition, the recombinant GhPEPC2 has been purified by expressing it in Escherichia coli and the catalytic properties of it were also investigated. The results showed that GhPEPC2 is a typical C3 PEPC with a higher Km (83.6 μM) and lower Vmax (8.0 μmol min 1 mg 1) compared with the C3 PEPCs previously reported.
Cloning and Ectopic Expression of ScYCF1 Gene from Saccharomyces cerevisiae in Cotton  [PDF]
Min Mu, Na Shu, Xuke Lu, Xiugui Chen, Shuai Wang, Junjuan Wang, Delong Wang, Weili Fan, Lixue Guo, Chao Chen, Wuwei Ye
Agricultural Sciences (AS) , 2018, DOI: 10.4236/as.2018.91005
Abstract: Yeast cadmium factor 1 (YCF1), is a member of the ATP-binding cassette (ABC) transporter family. To explore the functions of YCF1 of Saccharomyces cerevisiae (ScYCF1) in the cotton, ScYCF1 was cloned from Saccharomyces cerevisiae As2.375, with the full-length of 4548 bp. The bioinformatics analysis revealed that the largest component of ScYCF1 protein is leucine (12%). ScYCF1 is alkaline and positive charged, stable, and hydrophilic protein. The predictive secondary structure is mainly composed of α-helix areas, random coils and β-sheets. We constructed the pBI121-ScYCF1:GFP infusion expression vector and verified it by enzyme ingestion. The transient expression results of cotton pollen showed that the green fluorescence phenomenon of three kinds of upland cotton pollen significantly increased after transforming ScYCF1. The salt sensitive material upland cotton CCRI12 was transformed in vivo simultaneously, and the germination ability of trans-ScYCF1-gene T0 seeds was much better than the acceptor material CCRI12 under the stress of 100 mM NaCl saline solution. According to the gene nucleotide sequences, four pairs of primers were designed for molecular detection of T0 generation, and the sequencing results of PCR products of four specific primers evidence that the transgene is successful. Salt tolerance analysis of leaf discs of identified transgenic cotton showed that the chlorophyll content of leaf discs of transgenic cotton was higher than the content of the control cotton under salt stress. ScYCF1 gene was cloned and introduced into cotton, showing that ScYCF1 plays an important role in improving the salt tolerance of cotton.
The Core Binding Factor CBF Negatively Regulates Skeletal Muscle Terminal Differentiation  [PDF]
Ophélie Philipot,Véronique Joliot,Ouardia Ait-Mohamed,Céline Pellentz,Philippe Robin,Lauriane Fritsch,Slimane Ait-Si-Ali
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0009425
Abstract: Core Binding Factor or CBF is a transcription factor composed of two subunits, Runx1/AML-1 and CBF beta or CBFβ. CBF was originally described as a regulator of hematopoiesis.
短芒大麦dreb1基因的克隆与特性分析  [PDF]
王呈玉,张明哲,万生生,吕世友,马兰青,李彦舫
吉林农业大学学报 , 2006,
Abstract: ?在已知短芒大麦dreb1基因3'端序列的前提下,根据其他植物中dreb1转录因子基因序列,首次克隆了短芒大麦全长dreb1基因(899bp,具有1个编码220个氨基酸的完整阅读框)。序列分析和生物信息学分析表明,该基因具有转录因子的典型特征及ap2/erebp结构域,并且具有6个丝氨酸、2个苏氨酸、2个酪氨酸磷酸化位点,这可能与翻译后在冷胁迫抗性中发挥作用有关。
短芒大麦dreb1转录因子的克隆与特性分析  [PDF]
王呈玉?,万 甡?,翟凤艳?
西北农林科技大学学报(自然科学版) , 2009,
Abstract: [目的]克隆强抗寒性牧草--短芒大麦dreb1(dehydrationresponsiveelementbindingprotein1)转录因子,分析其生理生化特性,为理想抗逆工程基因的筛选和利用奠定理论基础.[方法]利用race-pcr(rapidamplificationofcdnaends-polymerasechainreaction)技术分离短芒大麦dreb1转录因子全长cdna序列,northern杂交和凝胶滞留试验分析其在逆境条件下的表达情况,及其与dre(dehydrationresponsiveelement)元件的结合活性.[结果]从强抗寒性短芒大麦中成功分离了1个新的dreb1类转录因子hbdreb1,该基因全长899bp,其蛋白序列中含有1个典型的ap2/erebpdna结构域及"pkk/rpagrxkfxetrhp"和"dsawr"、"lwsy"3个dreb1特征标签序列;序列比对分析表明,hbdreb1与其他植物的dreb1类转录因子的同源性较高.hbdreb1在转录水平上明显受冷胁迫诱导表达,具有结合dre-顺式作用元件的功能及作为转录因子必备的核定位特性.[结论]hbdreb1基因参与了非生物胁迫信号转导,具有提高植物抗寒性的潜能.
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