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Genomic Analysis of MicroRNA Promoters and Their Cis-Acting Elements in Soybean 被引量:5
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作者 LIU Yong-xin HAN Ying-peng +3 位作者 CHANG Wei ZOU Quan GUO Mao-zu LI Wen-bin 《Agricultural Sciences in China》 CSCD 2010年第11期1561-1570,共10页
MicroRNAs (miRNAs) are derived from distinct loci in the genome and play crucial roles in RNA-mediated gene silencing mechanisms that regulate cellular processes during development and stress responses of plants. Th... MicroRNAs (miRNAs) are derived from distinct loci in the genome and play crucial roles in RNA-mediated gene silencing mechanisms that regulate cellular processes during development and stress responses of plants. The miRNAs are approximately 21 nucleotides long and code for the complementary strand to a larger genic mRNA. They are often found within the complementary primary transcript (pri-miRNAs). In the past few years, a growing number of soybean miRNAs have been discovered, however, little is known about the transcriptional regulation of these miRNAs. In this study, promoters and cis-acting elements of soybean miRNAs were analyzed using the genomic data for the first time. A total of 82 miRNAs were located among 122 loci in genome, some were present as double or multiple copies. Five clusters that included ten miRNAs were found in genome, and only one cluster share the same promoter. A total of 191 promoters from 122 loci of the soybean miRNA sequences were found and further analyzed. The results indicated that the conserved soybean miRNA genes had a greater proportion of promoters than that of non-conserved ones, and the distribution of the transcript start sites (TSSs) and TATA-boxes found had different motif styles between conserved and non-conserved miRNA genes. Furthermore, the cis-acting elements 5' of the TSSs were analyzed to obtain potential function and spatiotemporal expression pattern of miRNAs. The data obtained here may lead to the identification of specific sequences upstream of pre-miRNAs and the functional annotation of miRNAs in soybean. 展开更多
关键词 MIRNAS SOYBEAN PROMOTER cis-acting elements miRNA clusters
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Isolation of cDNAs Enconding Two Distinct Transcription Factors Binding to DRE cis-acting Element Involved in Cold-and drought-induced Expression of Arabidopsis rd29A Gene 被引量:6
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作者 刘强 李进 +1 位作者 张贵友 赵南明 《Tsinghua Science and Technology》 SCIE EI CAS 1999年第3期1551-1559,共9页
Previous study has defined DRE (dehydration responsive element) cis acting element and its important role in expressions of Arabidopsis rd29A gene under cold, dehydration and high salt stresses. In order to... Previous study has defined DRE (dehydration responsive element) cis acting element and its important role in expressions of Arabidopsis rd29A gene under cold, dehydration and high salt stresses. In order to clarify the expression mechanism of rd29A gene, we isolated two cDNA clones that encoded DRE binding proteins ( DREB1 and DREB2 ) from cold and drought treated Arabidopsis plants, using DRE cis acting element in the promoter region of rd29A gene and yeast One Hybrid screening method. Experiments showed both DREB1 and DREB2 specifically interacted with DRE cis element. Homologous analysis showed no significant similarity between DREB1 and DREB2 in whole deduced amino acid sequences. However, both DREB1 and DREB2 proteins contained a conserved DNA binding domain (AP2/EREBP domain). Structural analysis of proteins also showed they had a nuclear localization signal (NLS) in their N terminal region and an acidic activation region in their C terminal region. AP2/EREBP domain is composed of 58 amino acids, which presents in a large family of plant genes encoding DNA binding proteins. We analyzed many plant transcription factors containing conserved AP2/EREBP domains. The 14th valine (V) and 19th glutamate (E) in the amino acid sequence of AP2/EREBP domains might be the consensus recognizing and binding to DRE cis element. Northern analysis indicated that DREB1 gene was induced by low temperature, whereas DREB2 gene was induced by dehydration and high salt stresses. Present studies suggest that the expression of rd29A gene under low temperature, dehydration and high salt stresses is regulated by DREB1 and DREB2 transcriptional factors in two separate signal transduction pathways, respectively. 展开更多
关键词 cis acting element transcription factor gene expression cDNA DNA\|binding domain
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Isolation and analysis of a TIR-specific promoter from poplar 被引量:12
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作者 Zheng Hui-quan Lin Shan-zhi Zhang Qian Zhang Zhen-zhen Zhang Zhi-yi Lei Yang Hou Lu 《Forestry Studies in China》 CAS 2007年第2期95-106,共12页
A 5' flanking region of the well-conserved Toll/interleukin-1 receptor domain (TIR)-encoding sequence was isolated from the genomic DNA ofMelampsora magnusiana Wagner resistant clones of hybrid triploid poplars [(... A 5' flanking region of the well-conserved Toll/interleukin-1 receptor domain (TIR)-encoding sequence was isolated from the genomic DNA ofMelampsora magnusiana Wagner resistant clones of hybrid triploid poplars [(Populus tomentosa × P. bolleana) × P. tomentosa]. Sequencing results and alignment analysis show that the obtained TIR-specific promoter (named as PtTIRp01) was 1,732 bp in length; moreover 3' region of the PtTIRp01 contains a responds to the 5' composition of TIR-NBS type gene PtDRG02, of 747 bp long 5' region of TIR-NBS type gene PtDRG02 and its 398 bp complete TIR-encoding sequence, which significantly corindicating that the obtained TIR-specific promoter region consists upstream region of promoter (985 bp). It was found that the 5' region of TIR-NBS type gene PtDRG02 was characterized in the downstream region of the transcriptional start, named as 5'-untranslated region (5' UTR), consisting of one 93 bp 5'-untranslation exon, one 213 bp intron and one 441 bp TIR-encoding open reading frame (ORF). In addition, several putative cis-acting motifs were present in the obtained TIR-specific promoter of PtDRG02, including one TATA box, one GC-rich, one AT-rich, one P-box, one 3-AF1 binding site, two CAAT boxes, two GT-1 motifs, three typical W-boxes, four I-boxes, and one multi-cis-acting fragment (MCF). The latter contains five types of regulatory elements (E4, G-box, ABRE motif, box 1 and HVA 1 s), most of which were homologous to the cis-acting regulatory elements involved in the activation of defense genes in plants. Thus, it can be suggested that TIR-specific promoter might be a pathogen-inducible promoter and be necessary for the inducible expression of defense-related genes. Key words Toll/interleukin- 1 receptor domain, promoter, Cis-acting element, poplar 展开更多
关键词 Toll/interleukin- 1 receptor domain PROMOTER cis-acting element POPLAR
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Cis-acting regulatory elements: from random screening to quantitative design 被引量:6
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《Frontiers of Electrical and Electronic Engineering in China》 CSCD 2015年第3期107-114,共8页
The cis-acting regulatory elements, e.g., promoters and ribosome binding sites (RBSs) with various desired properties, are building blocks widely used in synthetic biology for fine tuning gene expression. In the las... The cis-acting regulatory elements, e.g., promoters and ribosome binding sites (RBSs) with various desired properties, are building blocks widely used in synthetic biology for fine tuning gene expression. In the last decade, acquisition of a controllable regulatory element from a random library has been established and applied to control the protein expression and metabolic flux in different chassis cells. However, more rational strategies are still urgently needed to improve the efficiency and reduce the laborious screening and multifaceted characterizations. Building precise computational models that can predict the activity of regulatory elements and quantitatively design elements with desired strength have been demonstrated tremendous potentiality. Here, recent progress on construction of cis- acting regulatory element library and the quantitative predicting models for design of such elements are reviewed and discussed in detail. 展开更多
关键词 cis-acting regulatory element quantitative design synthetic biology random mutation modeling
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GCC box in Arabidopsis PDF1.2 promoter is an essential and sufficient cis-acting element in response to MeJA treatment 被引量:2
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作者 XIEBingyan LUXiangyang +1 位作者 YANGYuhong HUANGRongfeng 《Chinese Science Bulletin》 SCIE EI CAS 2004年第23期2476-2480,共5页
The expression of Arabidopsis PDF1.2 gene is regulated by jasmonic acid (JA) and ethylene (ET). It also has been well documented that GCC box is an element re- sponsive to ET,however, the responsive mechanism of JA in... The expression of Arabidopsis PDF1.2 gene is regulated by jasmonic acid (JA) and ethylene (ET). It also has been well documented that GCC box is an element re- sponsive to ET,however, the responsive mechanism of JA in such plant defense gene expression is unclear. In this paper, the authors define the essential cis-acting element in PDF1.2 promoter responsive to methyl jasmonate (MeJA) through fragment deletions and site-directed mutageneses combining Agrobacterium-mediated transient reporter gene expression in tobacco leaves. Firstly, the MeJA inducible expression of PDF1.2 was confirmed by using the upstream ?1.86 kb fragment of PDF1.2 gene. Secondly, the upstream –300— ?243 bp fragment of the promoter was evidenced to respond to MeJA. To further characterize this promoter region, three point mutations were introduced into the –300—?243 bp fragment of the promoter. This result showed that the muta- tion of GCC box abolished MeJA induction, whereas the mutations of the G box-like and the imperfect palindrome sequence did not significantly decrease MeJA inducible effect, indicating that GCC box in PDF1.2 is essential for MeJA induction. The sufficient responsiveness to MeJA of this GCC box was further investigated by 4×GCC fused up- stream to the CaMV 35S minimal promoter. This result sug- gested that the fused promoter was able to activate reporter gene expression in response to MeJA. Thus these results in- dicate that the GCC box in PDF1.2 is an essential and suffi- cient element to confer MeJA induction. 展开更多
关键词 PDF1.2基因 茉莉酮酸 乙烯 植物 甲基茉莉酮酸酯
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MicroRNA Primary Transcripts and Promoter Elements Analysis in Soybean (Glycine max L. Merril.) 被引量:1
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作者 LI Jing LIU Yong-xin +3 位作者 HAN Ying-peng LI Yong-guang GUO Mao-zu LI Wen-bin 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2013年第9期1522-1529,共8页
The importance of microRNA (miRNA) at the post-transcriptional regulation level has recently been recognized in both animals and plants. In recent years, many studies focused on miRNA target identification and funct... The importance of microRNA (miRNA) at the post-transcriptional regulation level has recently been recognized in both animals and plants. In recent years, many studies focused on miRNA target identification and functional analysis. However, little is known about the transcription and regulation of miRNAs themselves. In this study, the transcription start sites (TSSs) for 11 miRNA primary transcripts of soybean from 11 miRNA loci (of 50 loci tested) were cloned by a 5" rapid amplification of cDNA ends (5" RACE) procedure using total RNA from 30-d-old seedlings. The features consistent with a RNA polymerase II mechanism of transcription were found among these miRNA loci. A position weight matrix algorithm was used to identify conserved motifs in miRNA core promoter regions. A canonical TATA box motif was identified upstream of the major start site at 8 (76%) of the mapped miRNA loci. Several cis-acting elements were predicted in the 2 kb 5" to the TSSs. Potential spatial and temporal expression patterns of the miRNAs were found. The target genes for these miRNAs were also predicted and further elucidated for the potential function of the miRNAs. This research provides a molecular basis to explore regulatory mechanisms of miRNA expression, and a way to understand miRNA-mediated regulatory pathways and networks in soybean. 展开更多
关键词 SOYBEAN miRNA primary transcript RACE TATA box MOTIF PROMOTER cis-acting element
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A novel cold-inducible promoter,PThCAP from Tamarix hispida,confers cold tolerance in transgenic Arabidopsis thaliana
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作者 Xiaohong Guo Yandong Lv +4 位作者 Hongyu Li Nan Fu Guiping Zheng Lihua Liu Yuhua Li 《Journal of Forestry Research》 SCIE CAS CSCD 2018年第2期330-336,共7页
Our previous studies have revealed that the Th CAP gene plays a vital role in transgenic Populus(P.davidiana 9 P.bolleana) in response to cold stress.However,the regulatory mechanism of Th CAP gene expression has been... Our previous studies have revealed that the Th CAP gene plays a vital role in transgenic Populus(P.davidiana 9 P.bolleana) in response to cold stress.However,the regulatory mechanism of Th CAP gene expression has been unclear.In this study,the 50 flanking region of the Th CAP promoter(PTh CAP) was cloned using a genomewalking method.By analyzing cis-acting regulatory elements of PTh CAP,a DRE motif and MYC and MYB elements were found to be located in the promoter.To identify the regulatory elements that control the expression of the Th CAP gene promoter,a series of deletion derivatives ofPTh CAP,P1–P5,from the translation start code(-1538,-1190,-900,-718 and-375 bp),were fused to the GUS reporter gene,and then each deletion was stably introduced into Arabidopsis thaliana plants.Deletion analysis of the promoter suggested that only the P2 fragment had strong GUS expression in leaves and roots of A.thaliana exposed to low temperature stress.These results suggest that this290-bp region(-1190 to-900 bp),as an important part in PTh CAP,was associated with cold tolerance of A.thaliana.Our results provide evidence for the regulatory mechanism of Th CAP gene involved in the response to cold stress,and that the gene is promising candidate gene for genetic improvement of crops. 展开更多
关键词 Arabidopsis thaliana cis-acting elements GUS expression PThCAP ThCAP gene
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Functional analysis of cis-acting sequences regulating root-specific expression in transgenic tobacco 被引量:2
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作者 Lan Nan Huixin Lin +1 位作者 Yucheng Guan Fan Chen 《Chinese Science Bulletin》 SCIE EI CAS 2002年第17期1441-1445,共5页
Two different length fragments, RSF1 and RSF2 which contained the cis-acting sequences of root-specific gene TobRB7, were isolated from tobacco genome. The abilities of these fragments to direct root-specific expressi... Two different length fragments, RSF1 and RSF2 which contained the cis-acting sequences of root-specific gene TobRB7, were isolated from tobacco genome. The abilities of these fragments to direct root-specific expression were studied by fusing them to the β-glucuronidase (GUS) report gene with different directions. After the recombined vectors were transformed into tobacco, the expression pattern was performed by histochemical staining and the quantitative analysis of GUS activity. The data suggested that the cis-acting element of TobRB7 gene direct GUS expression not only as root-specific but also as bidirectional. In our studies, the short fragment, RSF2, performed stronger activity than RSF1 with any direction. The stronger activity of GUS expression was determined by reverse inserting of RSF1 or RSF2 than positive inserting. 展开更多
关键词 root TISSUE-SPEciFIC expression bidirectional PROMOTER cis-acting element TRANSGENIC plant.
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Developmental stage-specific factors in the mouse haematopoietic tissues binding to the 5'-flanking as-acting elements of humanε-globin gene 被引量:1
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作者 严志江 陈雅娣 钱若兰 《Chinese Science Bulletin》 SCIE EI CAS 1995年第9期778-783,共6页
The human ε-globin gene is expressed in a tissue-specific and developmental stage-specific manner. During the earliest stage of gestation, this gene is expressed in the yolksac, but is silenced completely at the 6th-... The human ε-globin gene is expressed in a tissue-specific and developmental stage-specific manner. During the earliest stage of gestation, this gene is expressed in the yolksac, but is silenced completely at the 6th-8th weeks of gestation. Recently, several studieson the transgenic mice have shown that the 5′-flanking DNA sequences of human 展开更多
关键词 HUMAN ε-globin gene cis-acting element trans-acting factor.
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Isolation and characterization of a novel cis-acting sequences regulating root-specific gene from Daucus carota L. 被引量:1
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作者 LIUYan GUOChang-an RENHaibo CHENFan 《Chinese Science Bulletin》 SCIE EI CAS 2004年第22期2393-2398,共6页
Aquaporins are ubiquitous channel proteins that facilitate the transport of water across cell membranes. Most of aquaporins are represent in more than one tissues, but some of them performed highest in roots. They are... Aquaporins are ubiquitous channel proteins that facilitate the transport of water across cell membranes. Most of aquaporins are represent in more than one tissues, but some of them performed highest in roots. They are be- lived to participate in water transport. DcRB7, a member of the aquaporin family, was isolated from somatic embryos of carrot and identified as a homologous gene of TobRB7. Further studies showed that the expression of DcRB7 was particular in carrot root. To investigate the transcription regulation of DcRB7, a 650-bp upstream sequence of DcRB7 was isolated by inverse PCR, and was fused to the β-glucuronidase (GUS) report gene. After the recombined vectors were transformed into tobacco, the expression pat- tern was performed by histochemical staining and the quan- titative analysis of GUS activity. The results indicated that the cis-acting element of DcRB7 gene directs GUS expression not only as root-specific but also as drought inducible. 展开更多
关键词 分离 表征 顺式作用元素 胡萝卜 基因调节 转基因作物 基因表达
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拟南芥RBCS-1A基因受光调节表达模式及其启动子遗传转化应用评价 被引量:7
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作者 习雨琳 周朋 +3 位作者 宋梅芳 李志勇 孟凡华 杨建平 《作物学报》 CAS CSCD 北大核心 2012年第9期1561-1569,共9页
RBCS编码光合碳同化关键酶核酮糖1,5-二磷酸羧化酶/加氧酶的小亚基,是控制植物光合作用的重要基因之一。本研究利用实时荧光定量PCR技术分析了拟南芥RBCS-1A受光调节的表达模式,结果表明,AtRBCS-1A表达受到光的诱导,同时具有组织表达特... RBCS编码光合碳同化关键酶核酮糖1,5-二磷酸羧化酶/加氧酶的小亚基,是控制植物光合作用的重要基因之一。本研究利用实时荧光定量PCR技术分析了拟南芥RBCS-1A受光调节的表达模式,结果表明,AtRBCS-1A表达受到光的诱导,同时具有组织表达特异性;运用生物信息学手段分析发现,该基因启动子序列中存在多个参与光应答的顺式作用元件;采用PCR技术从拟南芥基因组中分离到长度为1691bp的AtRBCS-1A启动子片段,将该片段与GUS报告基因融合构建植物表达载体并转化野生型拟南芥,对获得的转基因植株进行GUS染色,结果显示,AtRBCS-1A启动子是光诱导型和组织特异型启动子。以上结果初步证明,AtRBCS-1A启动子应用于植物遗传转化切实可行,具有重要应用价值。 展开更多
关键词 拟南芥 RBCS-1A基因 表达模式 光诱导表达 顺式作用元件
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Trametessp.AH28-2漆酶A的诱导合成及其基因5′-端调控区的克隆与分析 被引量:6
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作者 洪宇植 肖亚中 +4 位作者 房伟 张书祥 查向东 李剑凤 周宏敏 《生物工程学报》 CAS CSCD 北大核心 2005年第4期547-552,共6页
Trametessp.AH28_2漆酶同工酶的合成需要铜离子的存在,较高浓度的Cu2+有利于漆酶合成。在以葡萄糖为碳源补加0·5mmol/LCu2+的培养基中生长时,发酵液漆酶活性为44·3u/L,同时补加4·0mmol/L邻甲苯胺时,漆酶酶活提高到71... Trametessp.AH28_2漆酶同工酶的合成需要铜离子的存在,较高浓度的Cu2+有利于漆酶合成。在以葡萄糖为碳源补加0·5mmol/LCu2+的培养基中生长时,发酵液漆酶活性为44·3u/L,同时补加4·0mmol/L邻甲苯胺时,漆酶酶活提高到71·0u/L;而在补加Cu2+和邻甲苯胺的纤维二糖培养基中,酶活上升至2584u/L,为葡萄糖培养基的36·4倍。邻甲苯胺和铜离子诱导产生的漆酶同工酶组分,均为漆酶A(LacA)。竞争性RT_PCR分析表明,漆酶A基因(lacA)转录本的累积伴随有发酵液漆酶活性的增加,邻甲苯胺对lacA的调控发生在转录水平。lacA结构基因长2110bp,含有10个内含子;lacA的cDNA序列为1560bp,编码520aa的漆酶蛋白,其氨基酸序列与其它真菌漆酶具有较高的相似性。采用改进的反向PCR技术,扩增得到的lacA5′_端调控区长1881bp,分析表明,该区域上分布有1个TATA框、7个CAAT框和多个潜在的顺式作用元件序列位点,包括5个MRE元件、9个CreA结合位点、4个XRE元件、2个STRE元件和7个氮因子调控位点等。这些序列位点的存在部分地对应了菌株摇瓶发酵条件下lacA的表达规律。 展开更多
关键词 真菌漆酶 定量RT-PCR 长距离反向PCR 顺式作用元件
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小麦酪蛋白激酶TaCK2α启动子的克隆与启动活性分析
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作者 冯燕 吴志会 +5 位作者 曹雨欣 申松松 崔钟池 丁大烁 王海燕 刘大群 《河北农业大学学报》 北大核心 2025年第2期54-61,共8页
酪蛋白激酶2(Casein kinase 2,CK2)是在真核生物中普遍存在的组成型活性酶,已被证明广泛参与植物生长发育和环境胁迫反应。然而,CK2在小麦与叶锈菌互作过程中发挥的功能尚不清楚。本研究前期基于小麦TcLr19接种叶锈菌的转录组测序,筛选... 酪蛋白激酶2(Casein kinase 2,CK2)是在真核生物中普遍存在的组成型活性酶,已被证明广泛参与植物生长发育和环境胁迫反应。然而,CK2在小麦与叶锈菌互作过程中发挥的功能尚不清楚。本研究前期基于小麦TcLr19接种叶锈菌的转录组测序,筛选获得1个酪蛋白激酶基因TaCK2α,并证明其参与小麦抗叶锈病防御反应。为了进一步探究TaCK2α在小麦与叶锈菌互作过程中的转录调控机制,本研究基于中国春小麦数据库,获取TaCK2α基因上游启动子序列,克隆获得了TaCK2α启动子,命名为pTaCK2α。利用PlantCARE在线网站预测pTaCK2α包含的顺式作用元件,发现pTaCK2α序列含有启动子核心元件、逆境胁迫响应元件、激素响应元件、光响应元件;构建了pTaCK2α融合β-葡萄糖苷酸酶(β-Glucuronidase,GUS)、绿色荧光蛋白(Green fluorescent protein,GFP)报告基因的植物表达载体pTaCK2α-pBI121、pTaCK2α-pCamA,借助农杆菌转化本氏烟草,GUS组织化学染色及GFP荧光观察证明TaCK2α基因启动子具有启动活性。这些结果为探究TaCK2α在小麦抗叶锈病防御反应中的转录调控机制奠定了坚实的基础。 展开更多
关键词 小麦叶锈菌 酪蛋白激酶2α 顺式作用元件 启动子 启动活性
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金针菇L-赖氨酸合成通路基因鉴定及对不同光质的响应表达 被引量:11
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作者 陶永新 段静怡 +6 位作者 李依宁 李自燕 宋寒冰 张祺锶 黄嘉华 高玲玲 谢宝贵 《食用菌学报》 CSCD 北大核心 2018年第4期1-8,共8页
为了从分子水平探究提高金针菇赖氨酸含量的环境因子(温度、光照、湿度、CO_2)及其作用机理,本研究首先在金针菇(Flammulina filiformis)单核体L11基因组中鉴定L-赖氨酸合成通路基因,并预测其启动子区顺式元件。根据预测结果,采用红、... 为了从分子水平探究提高金针菇赖氨酸含量的环境因子(温度、光照、湿度、CO_2)及其作用机理,本研究首先在金针菇(Flammulina filiformis)单核体L11基因组中鉴定L-赖氨酸合成通路基因,并预测其启动子区顺式元件。根据预测结果,采用红、绿、蓝、红绿蓝复合4种光处理金针菇子实体,并分别用HPLC和定量PCR测定赖氨酸含量变化及其合成通路基因的响应表达。结果表明:金针菇赖氨酸由氨基己二酸(α-Aminoadipate,AAA)途径合成,通路中8步催化酶的编码基因及其序列在L11中全部得到鉴定。关键限速酶基因HCS、HAC1和SHD基因的启动子区存在G-box等光照响应元件。定量PCR表明,总体上看赖氨酸合成通路前7步的酶编码基因对4种光质均有响应表达。进一步比较本试验中4种不同光质的处理效果,发现蓝光照射后的金针菇子实体中,共有7个赖氨酸合成通路基因(HCS、HAC1、HAH、HDH、AAT、AAR和SR)表达量上调,同时赖氨酸含量最高。推测蓝光可能通过激活赖氨酸合成通路基因启动子区的G-box等顺式元件,诱导通路基因上调表达,进而提高了金针菇子实体中赖氨酸含量。 展开更多
关键词 食用真菌 赖氨酸生物合成 光照处理 顺式作用元件
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杂交构树UDP-葡萄糖脱氢酶基因编码蛋白的亚细胞定位及其启动子5'端缺失片段的功能分析 被引量:3
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作者 吉仁花 张文波 +4 位作者 林晓飞 包颖亮 特日格勒 包会嘎 白淑兰 《植物研究》 CAS CSCD 北大核心 2020年第6期932-942,共11页
为了研究杂交构树UDP-葡萄糖脱氢酶基因(DDBJ,BpUGDH基因登录号为LC457701)启动子不同区域的表达活性,利用5′端缺失及同源重组实验技术,将5个不同长度的BpUGDH启动子5′端缺失片段与GUS基因连接,并通过农杆菌介导法瞬时转化烟草;同时,... 为了研究杂交构树UDP-葡萄糖脱氢酶基因(DDBJ,BpUGDH基因登录号为LC457701)启动子不同区域的表达活性,利用5′端缺失及同源重组实验技术,将5个不同长度的BpUGDH启动子5′端缺失片段与GUS基因连接,并通过农杆菌介导法瞬时转化烟草;同时,为了定位BpUGDH基因编码的蛋白在细胞中表达的具体位置,利用GFP报告基因融合目的基因进行蛋白质的亚细胞定位。结果显示:BpUGDH基因启动子-244 bp以内的序列均能介导GUS基因的诱导表达,并且-973、-465、-355、-281和-244 bp之间的区域可能对BpUGDH基因启动子的活性发挥着至关重要的作用。另外,BpUGDH基因编码蛋白的亚细胞定位结果显示:BpUGDH位于叶绿体中。 展开更多
关键词 顺式作用元件 BpUGDH 亚细胞定位 杂交构树 5′端缺失分析法 瞬时表达
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猪嵌合RNA BCL2L2-PABPN1剪接调控、表达与亚细胞定位分析
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作者 杨秀芹 张倩 +4 位作者 李佳欣 郝婉君 张晓涵 何鑫淼 庞宇 《东北农业大学学报》 CAS CSCD 北大核心 2023年第1期79-87,共9页
相邻基因间顺式剪接(cis-splicing of adjacent gene,cis-SAGe)产生的嵌合RNA在细胞生长、疾病发生等生理活动中发挥重要作用,为探究cis-SAGe剪接调控机制,研究在前期克隆BCL2L2-PABPN1(BP)基础上,利用minigene、定点突变、瞬时转染、... 相邻基因间顺式剪接(cis-splicing of adjacent gene,cis-SAGe)产生的嵌合RNA在细胞生长、疾病发生等生理活动中发挥重要作用,为探究cis-SAGe剪接调控机制,研究在前期克隆BCL2L2-PABPN1(BP)基础上,利用minigene、定点突变、瞬时转染、半定量RT-PCR以及生物信息学分析等方法分析BP转录后剪接的调控元件,发现thyroid hormone receptor exon 9、gh-1 intron 3、srp40-exonic splicing enhancer、β-tropomyosin exon 6b等剪接增强子元件在嵌合子形成中发挥重要作用。同时比较分析BP与两个亲本基因组织表达以及亚细胞定位情况。研究结果将为进一步揭示BP转录后调控机制及功能提供基础。 展开更多
关键词 嵌合RNA BCL2L2-PABPN1 剪接 顺式调控元件
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拟南芥液泡H^(+)-ATP酶E1亚基基因AtTUF启动子的克隆及表达分析
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作者 孙海丽 梁静 +4 位作者 王文佳 丁位华 王妮娜 柳凯恒 李成伟 《华北农学报》 CSCD 北大核心 2022年第6期82-90,共9页
AtTUF基因编码液泡膜H^(+)-ATP酶的E1亚基,与跨液泡膜的物质运输密切相关。为了探明拟南芥AtTUF基因的组织表达特征及基因表达调控的分子机制,为AtTUF基因启动子的利用及AtTUF基因表达调控特征的深入研究提供理论依据,以哥伦比亚野生型... AtTUF基因编码液泡膜H^(+)-ATP酶的E1亚基,与跨液泡膜的物质运输密切相关。为了探明拟南芥AtTUF基因的组织表达特征及基因表达调控的分子机制,为AtTUF基因启动子的利用及AtTUF基因表达调控特征的深入研究提供理论依据,以哥伦比亚野生型拟南芥为材料,通过PCR扩增AtTUF的启动子(p AtTUF)序列,利用网站PlantCARE和PLACE预测其顺式作用元件,构建由其驱动半乳糖苷酶基因(Glucuronidase,GUS)的植物表达载体pBI121-p AtTUF,转化拟南芥获得转基因株系,通过GUS染色分析AtTUF启动子的组织表达模式及其对激素与逆境胁迫的响应特征,并对AtTUF在不同组织中的表达进行定量分析。结果表明,成功克隆到AtTUF基因开放阅读框上游2000 bp的启动子序列,其中含有多个响应光信号、脱落酸(ABA)、生长素、茉莉酸甲酯(MeJA)及干旱信号的顺式作用元件;转基因株系的GUS染色结果显示,AtTUF启动子驱动的GUS在拟南芥幼苗的叶与根中,成苗期的莲座叶、主茎、茎生叶、萼片、花瓣、花药、花粉粒、柱头、幼嫩角果及发育期种子中均有表达,而在幼苗期胚轴、成苗期的莲座叶侧枝、成熟期角果及种子中未检测到表达;且其表达受ABA、吲哚乙酸(IAA)、盐、暗处理及PEG的抑制,这与其含有多种光调控元件、激素调控元件及非生物胁迫响应元件相一致。实时荧光定量PCR结果显示,AtTUF在根、茎、叶、花、角果及种子中均有表达,尤以发育期角果中表达量最高,在花与干种子中表达量也较高,根中表达量最低。综上可知,AtTUF基因在植物各器官(尤其是生殖系统)的生长发育及响应ABA、IAA、盐、光及干旱信号的过程中均发挥着重要作用。 展开更多
关键词 拟南芥 AtTUF 启动子 顺式作用元件 组织表达
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陆地棉编码成花素同源基因GhFT1-A和GhFT1-D启动子的克隆及活性分析
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作者 蔡大润 李超 黄先忠 《石河子大学学报(自然科学版)》 CAS 2017年第3期305-311,共7页
在陆地棉全基因组中序列比对GhFT1基因在At和Dt基因组上的差异,设计特异性引物克隆其上游约1.8 kb的非编码序列作为目的启动子,通过测序结果区分At、Dt类型。本研究克隆出1701 kb GhFT1-A启动子和1771 kb GhFT1-D启动子(简称1.8 kb)。... 在陆地棉全基因组中序列比对GhFT1基因在At和Dt基因组上的差异,设计特异性引物克隆其上游约1.8 kb的非编码序列作为目的启动子,通过测序结果区分At、Dt类型。本研究克隆出1701 kb GhFT1-A启动子和1771 kb GhFT1-D启动子(简称1.8 kb)。序列比对分析发现了At和Dt基因组上不同GhFT1启动子间存在许多核苷酸序列差异。GhFT1-A和GhFT1-D启动子上存在大量的、作用广泛的顺式作用元件,并发现了一个顺式作用元件富集的重复片段区域。启动子活性检测证明了GhFT1基因上游1.8 kb的At和Dt启动子均具有活性,但At亚基因组启动子的活性弱于Dt型。陆地棉编码成花素同源基因GhFT1的启动子对于调控GhFT1-A和GhFT1-D的表达具有重要作用,并且它们之间的活性存在差异,暗示At和Dt基因组对开花调控作用不同。 展开更多
关键词 陆地棉 FLOWERING LOCUS T 1 成花素 启动子 顺式作用元件
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犬MC2R基因5′-UTR、3′-UTR序列测定、分析及其5′侧翼启动区序列分析
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作者 巴彩凤 范华 《辽宁医学院学报》 CAS 2008年第3期193-197,共5页
目的分离犬MC2R基因CDS部分序列和该基因5′、3′非翻译区,并对5′、3′非翻译区和5′侧翼的启动区进行分析。方法本研究采用反转录PCR(RT-PCR)和RNA连接酶介导的RACE(RLM-RACE)技术和BLAST分析软件。结果得到了5′、3′非翻译区和部分... 目的分离犬MC2R基因CDS部分序列和该基因5′、3′非翻译区,并对5′、3′非翻译区和5′侧翼的启动区进行分析。方法本研究采用反转录PCR(RT-PCR)和RNA连接酶介导的RACE(RLM-RACE)技术和BLAST分析软件。结果得到了5′、3′非翻译区和部分CDS片段的序列,他们的大小分别为168bp、1366bp和987bp,并预测了大约1500bp的5′侧翼的启动区域。结论对它们进行分析显示,该基因至少由两个外显子(exon1和exon2)组成,exon1和exon2的一部分编码5′非翻译区(5′-UTR),exon2其余的部分编码整个编码区。其启动区有inr,SF-1,SP1,CRE,PPRE,AP-1等多个顺式作用元件,这些为犬MC2R表达调控研究提供研究奠定基础。 展开更多
关键词 犬MC2R基因 基因结构 顺式作用元件 克隆
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人体β-珠蛋白基因5’旁侧调控元件与不同发育时期调控因子的相互作用
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作者 陈士友 蒋俶 钱若兰 《实验生物学报》 CSCD 1996年第4期379-384,共6页
人β-珠蛋白基因主要在成年期的骨髓中表达,在胎儿肝,成年肝和K 562细胞中则处于关闭状态。凝胶电泳阻抑法分析发现,在人胎儿肝,成年肝及K 562细胞的核蛋白抽提物中存在着不同的与β-珠蛋白基因5'旁侧调控元件(-372到-194 bp)相结... 人β-珠蛋白基因主要在成年期的骨髓中表达,在胎儿肝,成年肝和K 562细胞中则处于关闭状态。凝胶电泳阻抑法分析发现,在人胎儿肝,成年肝及K 562细胞的核蛋白抽提物中存在着不同的与β-珠蛋白基因5'旁侧调控元件(-372到-194 bp)相结合的红系组织特异性的调控因子。竟争试验的结果表明,成年肝和K 562细胞中的调控因子与β-珠蛋白基因5'旁侧调控元件相互作用的方式具有一定的相似性,这两种细胞的调控因子既结合于负调控区(NCR 2)也结合于正调控区,说明两种细胞中β-珠蛋白基因的关闭机制可能是相似的。胎儿肝的核蛋白因子只结合于负调控区,推测在胎儿期存在独特的关闭机制。 展开更多
关键词 细胞 Β-珠蛋白基因 调控元件 调控因子
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