PAP2(Production of anthocyanin pigment 2)编码MYB转录因子,与bHLH、WD40形成MYB-bHLH-WD40复合体来调控花青素合成。本研究从埃塞俄比亚芥和黑芥中分别克隆出2个和1个PAP2基因拷贝,分别命名为BcaB.PAP2、BcaC.PAP2、BniB.PAP2,这三...PAP2(Production of anthocyanin pigment 2)编码MYB转录因子,与bHLH、WD40形成MYB-bHLH-WD40复合体来调控花青素合成。本研究从埃塞俄比亚芥和黑芥中分别克隆出2个和1个PAP2基因拷贝,分别命名为BcaB.PAP2、BcaC.PAP2、BniB.PAP2,这三个基因均由3个外显子和2个内含子组成,均编码247个氨基酸。基于本研究克隆的PAP2基因序列和已报道的其他芸薹属PAP2基因序列,设计了3对能分别检测A、B、C基因组PAP2基因的引物,通过等位特异PCR可以区分芸薹属禹氏三角中的6个物种PAP2的基因组来源。选取芥菜型油菜B基因组的BjuB.PAP2基因构建过表达载体,转化拟南芥,其叶片变紫,表明PAP2基因正调控花青素合成。遮光处理10天后紫叶埃塞俄比亚芥的叶片紫色变淡,花青素含量下降了41.22%。荧光定量PCR和转录组研究表明,遮光处理植株叶片中BcaB.PAP2和BcaC.PAP2表达量均下降,花青素合成的结构基因查尔酮合成酶基因(CHS)、查尔酮异构酶基因(CHI)、黄烷酮3-羟化酶基因(F3H)、二氢黄酮醇还原酶基因(DFR)、花青素合成酶基因(ANS)、类黄酮3-葡糖基转移酶基因(UFGT)等基因的表达量也下降。本研究完成了对芸薹属植物中埃塞俄比亚芥和黑芥PAP2基因的克隆,并对其进行进化分析。根据本实验中克隆及其他已报道的PAP2基因序列设计等位基因特异性PCR引物,为芸薹属种间杂交后代PAP2基因的基因组传递鉴定提供了分子标记。通过BjuB.PAP2基因转拟南芥植株结果表明PAP2基因参与花青素的积累。紫叶埃塞俄比亚芥经遮光处理后,发现BcaPAP2基因及花青素合成结构基因的表达受光照的诱导。本研究在芸薹属中克隆PAP2基因,并初步验证PAP2的功能,为进一步解析芸薹属植物花青素合成的调控机制提供参考。展开更多
This paper reported firstly successful cloning of lycopene ε-cyclase （lbLCYe） gene from sweetpotato, lpomoea batatas （L.） Lam. Using rapid amplification of cDNA ends （RACE）, lbLCYe gene was cloned from sweetpot...This paper reported firstly successful cloning of lycopene ε-cyclase （lbLCYe） gene from sweetpotato, lpomoea batatas （L.） Lam. Using rapid amplification of cDNA ends （RACE）, lbLCYe gene was cloned from sweetpotato cv. Nongdafu 14 with high carotenoid content. The 1 805 bp cDNA sequence oflbLCYe gene contained a 1236 bp open reading frame （ORF） encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point （pI） of 6.95. IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD （flavinadenine dinucleotide）/NAD（P） （nicotinamide adenine dinucleotide phosphate）-binding domain, indicating that this protein shares the typical characteristics of LCYe proteins. The gDNA oflbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons. Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems. Transgenic tobacco （cv. Wisconsin 38） expressing [bLCYe gene accumulated significantly more ^-carotene compared to the untransformed control plants. These results showed that lbLCYe gene has an important function for the accumulation of carotenoids of sweetpotato.展开更多
Objective: To conduct the cloning identification and characterization of the sequence of human IL-17 A promoter so as to analyze the regulatory mechanism of the gene expression of IL-17. Methods: First of all, the pot...Objective: To conduct the cloning identification and characterization of the sequence of human IL-17 A promoter so as to analyze the regulatory mechanism of the gene expression of IL-17. Methods: First of all, the potential promoter region of IL-17 A was found by means of the bioinformatics methods. Then, it was cloned into the reporter vector with PCR technique. Finally, the activity of the test promoter was determined by dual luciferase reporter system. Results: Two transcriptional start points of the upper region, 600 bp and 1000 bp, of IL-17 A were obtained by PCR clone and proved to have certain activities by dual luciferase reporter system. Also, they could be activated by IL-17 A activator STAT3, which could start the expression of the reported gene. Conclusions: Clone established the regulatory region of human IL-17 A promoter, which provided bases to the subsequent function research.展开更多
Studies on the cold-responsive genes and cold signaling of woody species drop far behind in comparison to herbaceous plants.Due to similar lignified structure,perennial characteristic,and enhanced tolerance,it seems m...Studies on the cold-responsive genes and cold signaling of woody species drop far behind in comparison to herbaceous plants.Due to similar lignified structure,perennial characteristic,and enhanced tolerance,it seems much easier to find strongly antifreeze genes and obtain effective results in transgenic woody plants.In this study,Ammopiptanthus mongolicus,an evergreen,broadleaf and cold-resist leguminous shrub growing in the desert of Inner Mongolia,was used as a material for low-temperature induced gene isolation.Through differential expression analysis induced by low-temperature,thirteen up-regulated cDNAs were identified.One of them,AmEBP1,(accession number:DQ519359)confers enhanced cold-tolerance to both transgenic E.coli and transgenic Arabidopsis.Results suggest that AmEBP1 can stimulate the synthesis of ribosome and the dephosphyration of the α-subunit of initiation factor 2(eIF2α),and subsequently promote the translation process.By which the transgenic plants obtained increased cold-resistant ability.展开更多
As a major raw material for the textile industry and the most important fiber crop in the world,cotton is of great significance in Chinese economy.The development of cotton fiber can be divided
Eukaryotic translation initiation factor 1A(eIF1A)functions as an important regulatory factor of protein synthesis and plays a crucial role in responses to abiotic stresses in plants.However,little is known about the ...Eukaryotic translation initiation factor 1A(eIF1A)functions as an important regulatory factor of protein synthesis and plays a crucial role in responses to abiotic stresses in plants.However,little is known about the eIF1A gene involved in fruit development and stress response of mango.In this study,the MieIF1A-b gene was isolated from Mangifera indica,and contains a 435-bp open reading frame,which encodes a putative protein of 144 amino acids(GenBank accession number:KP676599).The predicted MieIF1A-b protein had a molecular weight of 16.39 kDa with a pI of 4.6.Sequence homology analysis showed that MieIF1A-b shared high homology with Elaeis guineensis,Manihot esculenta,and Populus trichocarpa,with 96 and 95%identity,respectively.Quantitative reverse transcriptative PCR(qRT-PCR)analyses indicated that MieIF1A-b was expressed in all tested tissues,and had the highest expression level in fruit 80 d after flowering.The expression of MieIF1A-b was obviously regulated by NaCl and H2O2 treatments in leaves.Functional analysis indicated that the overexpression of MieIF1A-b in transgenic Arabidopsis thaliana enhanced the growth,phenotype and salinity tolerance compared with wild-type(WT)plants.The results indicated that MieIF1A-b may be correlated with the control of fruit development and salt adaptation,and it was a candidate gene for abiotic stress in mango.展开更多
文摘PAP2(Production of anthocyanin pigment 2)编码MYB转录因子,与bHLH、WD40形成MYB-bHLH-WD40复合体来调控花青素合成。本研究从埃塞俄比亚芥和黑芥中分别克隆出2个和1个PAP2基因拷贝,分别命名为BcaB.PAP2、BcaC.PAP2、BniB.PAP2,这三个基因均由3个外显子和2个内含子组成,均编码247个氨基酸。基于本研究克隆的PAP2基因序列和已报道的其他芸薹属PAP2基因序列,设计了3对能分别检测A、B、C基因组PAP2基因的引物,通过等位特异PCR可以区分芸薹属禹氏三角中的6个物种PAP2的基因组来源。选取芥菜型油菜B基因组的BjuB.PAP2基因构建过表达载体,转化拟南芥,其叶片变紫,表明PAP2基因正调控花青素合成。遮光处理10天后紫叶埃塞俄比亚芥的叶片紫色变淡,花青素含量下降了41.22%。荧光定量PCR和转录组研究表明,遮光处理植株叶片中BcaB.PAP2和BcaC.PAP2表达量均下降,花青素合成的结构基因查尔酮合成酶基因(CHS)、查尔酮异构酶基因(CHI)、黄烷酮3-羟化酶基因(F3H)、二氢黄酮醇还原酶基因(DFR)、花青素合成酶基因(ANS)、类黄酮3-葡糖基转移酶基因(UFGT)等基因的表达量也下降。本研究完成了对芸薹属植物中埃塞俄比亚芥和黑芥PAP2基因的克隆,并对其进行进化分析。根据本实验中克隆及其他已报道的PAP2基因序列设计等位基因特异性PCR引物,为芸薹属种间杂交后代PAP2基因的基因组传递鉴定提供了分子标记。通过BjuB.PAP2基因转拟南芥植株结果表明PAP2基因参与花青素的积累。紫叶埃塞俄比亚芥经遮光处理后,发现BcaPAP2基因及花青素合成结构基因的表达受光照的诱导。本研究在芸薹属中克隆PAP2基因,并初步验证PAP2的功能,为进一步解析芸薹属植物花青素合成的调控机制提供参考。
基金supported by the China Agriculture Research System (Sweetpotato)the National High-Tech Research and Development Project of China(2011AA100607 and 2012AA101204)
文摘This paper reported firstly successful cloning of lycopene ε-cyclase （lbLCYe） gene from sweetpotato, lpomoea batatas （L.） Lam. Using rapid amplification of cDNA ends （RACE）, lbLCYe gene was cloned from sweetpotato cv. Nongdafu 14 with high carotenoid content. The 1 805 bp cDNA sequence oflbLCYe gene contained a 1236 bp open reading frame （ORF） encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point （pI） of 6.95. IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD （flavinadenine dinucleotide）/NAD（P） （nicotinamide adenine dinucleotide phosphate）-binding domain, indicating that this protein shares the typical characteristics of LCYe proteins. The gDNA oflbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons. Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems. Transgenic tobacco （cv. Wisconsin 38） expressing [bLCYe gene accumulated significantly more ^-carotene compared to the untransformed control plants. These results showed that lbLCYe gene has an important function for the accumulation of carotenoids of sweetpotato.
基金supported by the Natural Science Research Foundation of Anhui Provincial Education Department(KJ2016A464,KJ2016A478,KJ2014A161,KJ2015A248)General Natural Science Research Project of Anhui Education Department(KJ2015B022by,KJ2015B005by)+1 种基金the Foundation for Excellent Young Scholars of Anhui province(gxyq ZD2016163)the Scientific Research Innovation Team project of Anhui Colleges and Universities(2016-40)
文摘Objective: To conduct the cloning identification and characterization of the sequence of human IL-17 A promoter so as to analyze the regulatory mechanism of the gene expression of IL-17. Methods: First of all, the potential promoter region of IL-17 A was found by means of the bioinformatics methods. Then, it was cloned into the reporter vector with PCR technique. Finally, the activity of the test promoter was determined by dual luciferase reporter system. Results: Two transcriptional start points of the upper region, 600 bp and 1000 bp, of IL-17 A were obtained by PCR clone and proved to have certain activities by dual luciferase reporter system. Also, they could be activated by IL-17 A activator STAT3, which could start the expression of the reported gene. Conclusions: Clone established the regulatory region of human IL-17 A promoter, which provided bases to the subsequent function research.
文摘Studies on the cold-responsive genes and cold signaling of woody species drop far behind in comparison to herbaceous plants.Due to similar lignified structure,perennial characteristic,and enhanced tolerance,it seems much easier to find strongly antifreeze genes and obtain effective results in transgenic woody plants.In this study,Ammopiptanthus mongolicus,an evergreen,broadleaf and cold-resist leguminous shrub growing in the desert of Inner Mongolia,was used as a material for low-temperature induced gene isolation.Through differential expression analysis induced by low-temperature,thirteen up-regulated cDNAs were identified.One of them,AmEBP1,(accession number:DQ519359)confers enhanced cold-tolerance to both transgenic E.coli and transgenic Arabidopsis.Results suggest that AmEBP1 can stimulate the synthesis of ribosome and the dephosphyration of the α-subunit of initiation factor 2(eIF2α),and subsequently promote the translation process.By which the transgenic plants obtained increased cold-resistant ability.
基金This work was funded by grants fromthe National Basic Research and Development Program(2004CB117304)the Hi-tech Research and Development Program of China (2007AA10Z115)
文摘As a major raw material for the textile industry and the most important fiber crop in the world,cotton is of great significance in Chinese economy.The development of cotton fiber can be divided
基金supported by the National Natural Science Foundation of China (31660561)the Natural Science Foundation of Guangxi, China (2015GXNSFAA139052)+3 种基金the Key Research and Development Project of Guangxi, China (GXKJ-AB17292010)the Major Science and Technology Projects of Guangxi, China (GXKJ-AA17204097-3 and GXKJ-AA172040 26-2)the Innovation Team of Guangxi Mango Industry Project, China (nycytxgxcxtd-06-02)the State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, China (SKLCUSA-a201906)
文摘Eukaryotic translation initiation factor 1A(eIF1A)functions as an important regulatory factor of protein synthesis and plays a crucial role in responses to abiotic stresses in plants.However,little is known about the eIF1A gene involved in fruit development and stress response of mango.In this study,the MieIF1A-b gene was isolated from Mangifera indica,and contains a 435-bp open reading frame,which encodes a putative protein of 144 amino acids(GenBank accession number:KP676599).The predicted MieIF1A-b protein had a molecular weight of 16.39 kDa with a pI of 4.6.Sequence homology analysis showed that MieIF1A-b shared high homology with Elaeis guineensis,Manihot esculenta,and Populus trichocarpa,with 96 and 95%identity,respectively.Quantitative reverse transcriptative PCR(qRT-PCR)analyses indicated that MieIF1A-b was expressed in all tested tissues,and had the highest expression level in fruit 80 d after flowering.The expression of MieIF1A-b was obviously regulated by NaCl and H2O2 treatments in leaves.Functional analysis indicated that the overexpression of MieIF1A-b in transgenic Arabidopsis thaliana enhanced the growth,phenotype and salinity tolerance compared with wild-type(WT)plants.The results indicated that MieIF1A-b may be correlated with the control of fruit development and salt adaptation,and it was a candidate gene for abiotic stress in mango.
