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Effects of Light Stress on Oxygen Evolution and Photochemical Energy Stor age of Hybrid Poplar Clones Determined by Photoacoustic Technique 被引量:1
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作者 张守仁 高荣孚 《Acta Botanica Sinica》 CSCD 2000年第8期818-823,共6页
The oxygen evolution, thermal dissipation, and photochemical energy storage of three hybrid poplar clones, namely the triploid clone B342, the diploid clone B11 [(Populus alba×P. glandulosa)×(P.tomentosa... The oxygen evolution, thermal dissipation, and photochemical energy storage of three hybrid poplar clones, namely the triploid clone B342, the diploid clone B11 [(Populus alba×P. glandulosa)×(P.tomentosa×P.bolleana)], and the triploid clone B346 [(P.tomentosa×P. bolleana)×(P. alba×P.glandulosa)], under light stress were studied using photoacoustics. The oxygen evolution signal and photochemical energy storage varied negatively with the pretreatment_PFD (photon flux density), whereas the thermal signal varied positively with the pretreatment_PFD. Photochemical energy storage was reallocated to PSⅡ more than to PSⅠ, while the photochemical energy storage in PSⅠ was more stable than that in PSⅡ when subjected to light stress. The inhibitors streptomycin (SM), dithiothreitol (DTT) and sodium fluoride (NaF) could all affect the oxygen evolution signal. Clones B11 and B342 were more resistant to light stress than clone B346. 展开更多
关键词 hybrid poplar clones light stress oxygen evolution signal photochemical energy storage PHOTOACOUSTICS INHIBITORS
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川黔紫薇LeAGL11基因克隆表达分析及转录自激活检测
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作者 李雪露 陆柳淑 +5 位作者 邓涪元 李露 雷宇行 彭继庆 何钢 乔中全 《中南林业科技大学学报》 CAS CSCD 北大核心 2024年第6期197-206,共10页
【目的】探究转录因子AGL11在紫薇属紫薇自交及川黔紫薇与紫薇杂交过程中不同时间的差异性表达及其在酵母中的转录自激活特性,为研究紫薇远缘杂交结实率低的分子机制奠定基础。【方法】以‘紫韵’紫薇自交授粉和 ‘紫韵’紫薇ב川... 【目的】探究转录因子AGL11在紫薇属紫薇自交及川黔紫薇与紫薇杂交过程中不同时间的差异性表达及其在酵母中的转录自激活特性,为研究紫薇远缘杂交结实率低的分子机制奠定基础。【方法】以‘紫韵’紫薇自交授粉和 ‘紫韵’紫薇ב川黔1号’川黔紫薇杂交授粉后 24、48 和72 h的雌蕊为材料,克隆LeAGL11基因,通过生物信息学分析其理化性质等,采用实时荧光定量方法分析该基因在不同授粉方式和不同阶段的相对表达量,通过DNA重组技术构建LeAGL11的酵母表达载体,转化至Y2HGold感受态细胞内进行转录自激活检测。【结果】从川黔紫薇中克隆得到LeAGL11基因,该基因的编码序列长666 bp,编码221个氨基酸,LeAGL11蛋白无信号肽和跨膜结构域,不属于膜蛋白,属于亲水性蛋白。氨基酸序列比对和进化树分析显示其与紫薇、石榴和葡萄的AGL11蛋白都有较高的相似度;蛋白质结构预测和序列分析表明其具有MADS-box基因家族中典型的MADS-box和K-box保守结构域。实时荧光定量试验分析不同阶段的相对表达量表明LeAGL11基因在杂交后呈现出上调的趋势,在自交后呈现出先下调后上调的趋势,在自交授粉24 h时相对表达量最高,自激活检测发现pGBKT7-LeAGL11重组质粒在缺色氨酸的培养基中生长,在SD/-Trp+AbA+X-α-gal培养基中没有发生颜色变化。【结论】LeAGL11基因在紫薇自交过程及川黔紫薇和紫薇杂交过程中的相对表达量具有一定的差异性,且LeAGL11无转录自激活活性,可用于后续试验。 展开更多
关键词 紫薇属 远缘杂交 AGL11 基因克隆 基因表达
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杂交兰ChCAO基因克隆及其在叶艺品系中的表达特征
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作者 林榕燕 孔兰 +2 位作者 吴建设 林兵 钟淮钦 《西北植物学报》 CAS CSCD 北大核心 2024年第3期421-429,共9页
【目的】叶绿素酸酯a加氧酶(CAO)是叶绿素b形成过程中的关键酶,对杂交兰CAO基因进行克隆及表达特征分析可为探究其在杂交兰叶艺形成中的调控作用奠定基础。【方法】以杂交兰‘紫妍氏’(K21)及其叶艺品系‘中透紫妍氏’(K21-3)为试验材料... 【目的】叶绿素酸酯a加氧酶(CAO)是叶绿素b形成过程中的关键酶,对杂交兰CAO基因进行克隆及表达特征分析可为探究其在杂交兰叶艺形成中的调控作用奠定基础。【方法】以杂交兰‘紫妍氏’(K21)及其叶艺品系‘中透紫妍氏’(K21-3)为试验材料,用RT-PCR和RACE技术从叶片中克隆获得ChCAO基因,对ChCAO进行结构特征、理化性质、序列比对以及系统进化关系等分析;用qRT-PCR法对ChCAO在不同组织及叶艺品系叶片中的表达特性进行分析;并用VIGS技术对ChCAO进行沉默表达。【结果】ChCAO基因编码区长1608 bp,编码535个氨基酸,ChCAO与墨兰CAO亲缘关系最近,并与其他兰科植物CAO聚为一类。qRT-PCR结果显示,ChCAO基因表达具有组织特异性,在叶中相对表达量最高,根中相对表达量最低;此外,ChCAO在K21绿叶和K21-3绿叶区域叶片中的相对表达量显著高于K21-3叶艺区域叶片中的相对表达量。构建该基因的VIGS沉默载体转化烟草,发现沉默ChCAO后烟草叶片呈黄化状态,叶片中的叶绿素含量及ChCAO基因的相对表达量也显著降低。【结论】克隆获得了杂交兰ChCAO基因,并对其功能进行了初步鉴定,为进一步研究杂交兰叶艺形成机理提供重要依据。 展开更多
关键词 杂交兰 ChCAO基因 基因克隆 表达分析
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Growth variations and stability analyses of seven poplar clones at three sites in northeast China 被引量:2
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作者 Xiaona Pei Luping Jiang +4 位作者 Ammar khalil Mohamed Ahmed Hongying Yu Rizheng Chong Xiangling You Xiyang Zhao 《Journal of Forestry Research》 SCIE CAS CSCD 2021年第4期1673-1680,共8页
Growth characteristics have complex inheritance patterns and genotype(G) by environment(E) interaction make predicting tree response to environmental changes difficult.In this study,the growth of seven poplar clones a... Growth characteristics have complex inheritance patterns and genotype(G) by environment(E) interaction make predicting tree response to environmental changes difficult.In this study,the growth of seven poplar clones at three different sites was taken as the research focus,and heights and basal diameters were investigated in the second growing season.An ANOVA showed that all main effects,site,clone number and their interactions were highly significant in the overall F-tests.The coefficients of variation and repeatability of different traits ranged from 15.5 to 43.9%and from 0.549 to 0.912,respectively.AMMI(Additive Main Effects and Multiplicative Interaction) analysis results showed that genotype,environment and G × E interaction were significantly highly correlated.The stability analysis indicated that different clones showed different growth traits on different sites,which suggests that elite clones should be selected separately for different sites.Based on the growth traits,under a 10% selection rate,three clones were selected for different sites and the genetic gains of growth traits ranged from 4.7 to 11.2%.The three selected clones could be used to establish plantations in the future in different sites. 展开更多
关键词 POPLAR hybrid clone GENOTYPE Environmental interaction Stability REPEATABILITY
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Genetic variation of height growth rhythm between clones of Larix kaempferi × L. gmelini based on logistic models 被引量:1
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作者 Chunming Li Hui Xia +4 位作者 Hui Bai Hongmei Wang Yajuan Xing Xiyang Zhao Xiaomei Sun 《Journal of Forestry Research》 SCIE CAS CSCD 2018年第5期1387-1394,共8页
Fifty-three larch interspecific hybrid clones(Larix kaempferi × L.gmelini) and their parent clones were used for growth curve analysis of height variations.The growth curves of the 55 clones were 'S'-shaped a... Fifty-three larch interspecific hybrid clones(Larix kaempferi × L.gmelini) and their parent clones were used for growth curve analysis of height variations.The growth curves of the 55 clones were 'S'-shaped and 36 exhibited similar curves as the male parent.The coefficients of the logistic models were higher than 0.943,indicating that our results were effective in the simulation of the growth curves.ANOVA analysis showed significant differences in height of different clones (P/0.01).Average date of maximum height growth was Day 173,and average duration of rapid growth lasted for 50 days.Annual average increase in height was 9.7cm d^(-1) and daily average increase was 0.2 cm.The ratio of GR to the total annual increase in height ranged from 51.2 to 68.8%,with the average being 59.8%.There was a positive correlation between k values and plant heights which benefited from the evaluation of early plant height.There was also a positive correlation between GR(growth stage),GD(plant height) and annual increase in height.These results are informative to the evaluation of the elite clone selection and provide a theoretical basis for breeding and management. 展开更多
关键词 Larix kaempferi ×L. gmelini hybrid clones Logistic modeling Plant height variation
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白菜种子cDNA酵母文库的构建及BrTTG1互作蛋白的筛选及分析
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作者 任延靖 张鲁刚 +2 位作者 赵孟良 李江 邵登魁 《生物技术通报》 CAS CSCD 北大核心 2024年第2期223-232,共10页
【目的】通过构建白菜种子的cDNA文库,筛选WDR 40蛋白TRANSPARENT TESTA GLABRA 1(TTG1)的互作蛋白,探究TTG1参与MBW三元复合体调控种皮原花青素形成的分子机制。【方法】以棕籽白菜自交系‘B147’的种子为材料,提取总RNA并建立cDNA文库... 【目的】通过构建白菜种子的cDNA文库,筛选WDR 40蛋白TRANSPARENT TESTA GLABRA 1(TTG1)的互作蛋白,探究TTG1参与MBW三元复合体调控种皮原花青素形成的分子机制。【方法】以棕籽白菜自交系‘B147’的种子为材料,提取总RNA并建立cDNA文库,通过gateway技术构建诱饵载体pGBKT7-TTG1并进行酵母双杂交筛库。【结果】酵母文库库容为1.2×10^(7)CFU,文库滴度是5.0×10^(7)CFU/mL,插入片段平均长度大于1000 bp,诱饵载体在酵母中无自激活活性。通过构建的诱饵载体pGBKT7-TTG1与构建的cDNA文库杂交,共获得了38个阳性互作蛋白,功能预测显示其中一个蛋白注释为MYB转录因子,注释为MYB73,序列分析结果显示该基因含有R2R3-MYB型抑制子保守基序C1和C2,推测该基因为白菜中参与种皮颜色形成的R2R3-MYB型抑制子,暗示着白菜中可能存在不同MYB转录因子参与的调控网络,影响着原花青素的形成。【结论】本研究构建了白菜种子组织的酵母双杂交cDNA文库,获得了38个TTG1阳性互作蛋白,首次挖掘到了可能影响白菜种皮颜色原花青素形成的R2R3-MYB型抑制子MYB73,为后期探究白菜种皮原花青素的调控网络奠定良好的基础。 展开更多
关键词 白菜种皮颜色 CDNA文库 酵母双杂交 互作蛋白 MYB73 基因克隆 表达分析
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钩藤UrLAMT基因及其启动子的克隆与分析
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作者 李永权 刘淼 +4 位作者 上官黎阳 王晓红 胡涛 唐柳 张明生 《西北农林科技大学学报(自然科学版)》 CSCD 北大核心 2024年第2期145-154,共10页
【目的】克隆钩藤(Uncaria rhynchophylla)中马钱苷酸甲基转移酶(loganic acid methyltransferase,LAMT)基因及其启动子,对UrLAMT基因的组织表达及其对生物和非生物胁迫的响应进行分析,并对UrLAMT及其启动子的生物信息学进行分析,为进... 【目的】克隆钩藤(Uncaria rhynchophylla)中马钱苷酸甲基转移酶(loganic acid methyltransferase,LAMT)基因及其启动子,对UrLAMT基因的组织表达及其对生物和非生物胁迫的响应进行分析,并对UrLAMT及其启动子的生物信息学进行分析,为进一步研究UrLAMT转录调控奠定基础。【方法】基于钩藤的转录组数据设计引物,采用PCR从钩藤cDNA中克隆UrLAMT序列,并对其进行生物信息学分析;采用实时荧光定量PCR,分析UrLAMT在钩藤不同组织(根、茎、叶、花、果实、钩)中的表达,以及对外源茉莉酸甲酯(MeJA)、遮光/复光胁迫的响应;利用FPNI-PCR技术克隆UrLAMT上游启动子序列,并验证其活性,结合酵母单杂交试验,分析相应转录因子与UrLAMT启动子的调控关系。【结果】克隆得到钩藤UrLAMT序列,其长度为1137 bp,共编码378个氨基酸。UrLAMT蛋白质相对分子质量为42.64 ku,理论等电点为5.76,为亲水性蛋白,无信号肽,不含跨膜结构,定位在细胞质中;UrLAMT基因在钩藤叶中表达量最高,其次为根;UrLAMT均能响应MeJA和光;其与短小蛇根草和长春花的进化关系较近。UrLAMT启动子序列长度为1141 bp,除核心响应元件外还含有多个光响应元件,UrLAMT启动子在酵母中与光调控转录因子HY5存在相互作用。【结论】获得了UrLAMT基因及其启动子序列,其在钩藤叶中表达量最高,可能受光诱导。 展开更多
关键词 钩藤 马钱苷酸甲基转移酶 基因克隆 启动子克隆 酵母单杂交
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天香百合、药百合黄酮醇合成酶FLS基因克隆和表达分析
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作者 刘筱琳 孙婷婷 +1 位作者 杨捷 何恒斌 《浙江农业学报》 CSCD 北大核心 2024年第2期344-357,共14页
以天香百合(Lilium auratum)和药百合(L.speciosum var.gloriosoides)为研究材料,分别克隆获得黄酮醇合成酶(flavonol synthase, FLS)基因,命名为LaFLS和LsFLS。实验结果表明,LaFLS和LsFLS基因均含完整的开放阅读框1 035 bp,均编码344... 以天香百合(Lilium auratum)和药百合(L.speciosum var.gloriosoides)为研究材料,分别克隆获得黄酮醇合成酶(flavonol synthase, FLS)基因,命名为LaFLS和LsFLS。实验结果表明,LaFLS和LsFLS基因均含完整的开放阅读框1 035 bp,均编码344个氨基酸,氨基酸序列高度保守,均具有DIOX-N结构域和2-酮戊二酸和铁(Ⅱ)依赖性双加氧酶结构域,属于2-酮戊二酸和铁(Ⅱ)依赖性双加氧酶超家族;系统进化分析表明,LaFLS和LsFLS除与东方系百合西伯利亚和索邦的FLS亲缘关系最近外,与百合科郁金香(Tulipa fosteriana)等亲缘关系较近;生物信息学分析显示,LaFLS和LsFLS蛋白无信号肽序列和跨膜结构域,均为亲水性蛋白,亚细胞定位结果显示二者主要定位在细胞质中。基因表达分析结果表明,在花蕾发育过程中,LaFLS和LsFLS随花蕾发育出现先上升后下降再上升的趋势,而且在花被片无色区的表达量显著高于有色区域。 展开更多
关键词 东方百合 黄酮醇合成酶基因 黄酮醇 克隆 基因表达
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Physical Location of the Rice Pi-5(t), Glh and RTSV Genes by ISH of BAC Clones 被引量:24
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作者 Yan Huimin, Song Yunchun, Li Lijia, Bi Xuezhi, Fu Binying(College of Life Sciences, Wuhan University, Wuhan 430072, China) 《Wuhan University Journal of Natural Sciences》 EI CAS 1998年第2期226-230,共5页
Mapping of low or single-copy sequences on plant chromosomes has proven difficult because of very low frequency of signal detection. Rice BAC library is being used widely in rice genome research due to its distinctive... Mapping of low or single-copy sequences on plant chromosomes has proven difficult because of very low frequency of signal detection. Rice BAC library is being used widely in rice genome research due to its distinctive advantages over other library systems. In this study, two biotin-labeled rice BAC clones closely linked to a rice blast resistance, green leafhopper resistance and tungro spherical virus resistance gene,Pi-5(t), Glh, RTSV, werein situ hybridized to rice chromosomes. They were located on the long arm and short arm of chromosome 4 with FL value of 40%and 100%respectively. The frequency of signal detection reached 46.8%and 59.2%. The signal location were consistent with the selective marker on rice saturated molecular map. The results demonstrated the advantages to locate BAC clones to chromosomes byin situ hybridization and will facilitate the rice low or single-copy gene location by using the BAC library. 展开更多
关键词 Key words BAC clone biotin labeling in situ hybridization
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Identification of differentially expressed genes associated with bud dormancy release in tree peony(Paeonia suffruticosa) by suppression subtractive hybridization 被引量:2
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作者 HUANG Xin ZHENG Guo-sheng +1 位作者 DAI Si-lan GAI Shu-peng 《Forestry Studies in China》 CAS 2008年第2期88-94,共7页
A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies. In order to identify genes that are highly expressed in buds released from dormancy, 588 clones were examin... A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies. In order to identify genes that are highly expressed in buds released from dormancy, 588 clones were examined by differential screening. Of these, 185 clones were selected to be sequenced. A total of 37 unique sequences were obtained of which only 31 sequences have matches in the NCBI database or the Arabidopsis thaliana protein database. Semi-quantitative RT-PCR was used to confirm further the expression profiles for 12 transcripts identified within the subtractive cDNA library. Gene ontology analyses indicated that many of the different genes identified have unknown or hypothetical functions while it is speculated that other genes play different mo- lecular roles. In our study, genes involved in bud dormancy release were growth-related or stress-responsive, while low-temperature-induced ribosomal proteins may also play a role in bud dormancy release. Our results provide interesting information for further understanding of the molecular mechanism of bud dormancy release in tree peonies. 展开更多
关键词 cDNA clone DORMANCY subtractive hybridization tree peony (Paeonia suffruticosa)
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Clones No 1333 of Populus alba×P.davidiana and No 1132 of P.davidiana×P.alba×P.daviana
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作者 王继军 刘玉纯 +2 位作者 张方春 陆国君 彭勇军 《Journal of Forestry Research》 SCIE CAS CSCD 1997年第2期74-76,共3页
The hybridization experiment was initiated in 1975, in which the parents of P davhaana were collectedfrom Dailing, Heilongiiang Province, P. suaveolens were from Baicheng, P. simonii from Zhaodong of Heilongjiang Prov... The hybridization experiment was initiated in 1975, in which the parents of P davhaana were collectedfrom Dailing, Heilongiiang Province, P. suaveolens were from Baicheng, P. simonii from Zhaodong of Heilongjiang Prov-ince, and P. tremula from Shanxi Province. Clones No 1333 of P. Alba x P. davidiana and No 1132 of P davidiana x (Palba x P. davidiana F1) had greater genetic variation and heritability in clones tested. 展开更多
关键词 clones hybridization ASPEN
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Screening and identification of binding proteins to interferon-α from a cDNA library by yeast-two hybrid system
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作者 JIAN HUI QU JUN CHENG +7 位作者 LING XIA ZHANG YAN WEI ZHONG YUAN YANG JIANG GUO LI YING ZHANG YAN LIU LI WANG JIU ZENG DAI 《Journal of Microbiology and Immunology》 2005年第3期187-192,共6页
The aim of this study is to screen proteins interacting with interferon-α (IFN-α). The IFN-α gene was amplified by polymerase chain reaction (PCR) and cloned into pGBKT-/vector, then the resulted pGBKT-/-IFN-α... The aim of this study is to screen proteins interacting with interferon-α (IFN-α). The IFN-α gene was amplified by polymerase chain reaction (PCR) and cloned into pGBKT-/vector, then the resulted pGBKT-/-IFN-α vector was transformed into yeast strain AH109. The transformed yeast AH109 was mated with yeast Y187 containing liver cDNA library plasmid in 2 × YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade and SD/-Trp-Leu-His) con- taining X-α-gal for selection. After plasmid extraction and enzyme cutting analysis, the blue colonies were subjected to sequence analysis and the results were analyzed by bioinformatics. The results showed that IFN-α was successful cloned into the pGBKT7 vector. IFN-α was expressed and there was no selfactivation and toxicity in AH109. Thirty-four positive colonies were obtained after yeast-two hybrid technique screening. After sequence analysis, eight clones were found to have a binding effect with IFN-α protein. IFN-α was successfully cloned into the pGBKT7 vector. IFN-α protein was expressed and there was no self-activation and toxicity in AH109. Eight proteins that interacted with IFN-α, including vitronectin, fibrinogen A alpha polypeptide, HIV-1 Tat interactive protein 2, arginase, NADH dehydrogenase 1 beta subcomplex, transferrin receptor 2 alpha (TFR2), HCC-1, alcohol dehydrogenase IB (ADHIB) have been identified as IFN-α-binding proteins. 展开更多
关键词 IFN-α Yeast-two hybrid system Gene cloning
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Improved methods for cloning and detection in the yeast two hybrid assay
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作者 Shenshen Zou Qianyu Wang +5 位作者 Xin Kang Yilie Liao Yong Chen Yutao Liu Gaoyi Min Yongheng Liang 《Advances in Bioscience and Biotechnology》 2012年第7期928-935,共8页
The yeast two-hybrid (Y2H) mating assay is a powerful method for detecting protein-protein interactions. Firstly, the gene of interest is cloned into specific Y2H vectors. Although multiple innovations in cloning meth... The yeast two-hybrid (Y2H) mating assay is a powerful method for detecting protein-protein interactions. Firstly, the gene of interest is cloned into specific Y2H vectors. Although multiple innovations in cloning methods were made in the past two decades, the conventional cloning method of restriction-enzyme (RE) digestion followed by ligation is still widely used. Unfortunately, many researchers, especially new-comers, often encounter difficulties in cloning a gene into a desired vector. Secondly, interaction between two proteins is commonly detected by growth of the diploids in specific media. This step takes about two weeks. Here, we describe improved cloning and detection procedures for the Y2H assay that accelerate the research progress. The changes in procedures involve running an agarose gel after the doubly digested vector and insert are ligated in the cloning step to determine the efficiency of RE digestion and ligation, and performing an additional replica-plating on plates for earlier assessment of interaction in the detection step. We show an example of Y2H interaction between Trs23 and Trs120 (respective subunits of TRAPP I and TRAPP II), as a proof of concept. By following the improved methods described here, the chances of successful cloning increased and the time for the whole Y2H experimental process is significantly shorter. 展开更多
关键词 CONVENTIONAL METHOD Improved METHOD CLONING YEAST Two hybrid PROTEIN-PROTEIN Interactions
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兰州鲇NPY基因的克隆和表达分析 被引量:1
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作者 赖章龙 阮超岭 +11 位作者 肖伟 刘彦斌 俞兆曦 刘凯 赛清云 田永华 张锋 杨瑞兰 柳婷 杨立强 王玉涛 连总强 《华北农学报》 CSCD 北大核心 2023年第5期227-238,共12页
针对兰州鲇人工驯食转化率、苗种培育成活率低等问题,为利用基因编辑技术创制和培育易驯食兰州鲇新种质,开展了兰州鲇神经肽Y基因(NPY)的克隆及其表达分析研究。结果表明:兰州鲇NPY基因cDNA序列全长为775 bp,包括288 bp的开放阅读框,编... 针对兰州鲇人工驯食转化率、苗种培育成活率低等问题,为利用基因编辑技术创制和培育易驯食兰州鲇新种质,开展了兰州鲇神经肽Y基因(NPY)的克隆及其表达分析研究。结果表明:兰州鲇NPY基因cDNA序列全长为775 bp,包括288 bp的开放阅读框,编码95个氨基酸,含有1个跨膜螺旋和1个信号肽胰腺激素类识别结合位点。兰州鲇NPY蛋白序列与南方鲇的一致性最高,为98.95%。荧光定量PCR检测发现NPY mRNA在脑组织中表达量最高,其次是下丘脑,肌肉中表达量最低。不同年龄雄性各组织中NPY mRNA表达水平均高于雌性;且雌雄性别组间均表现出随年龄的增长NPY mRNA表达下降的特征。利用原位杂交技术进行脑组织细胞定位发现,脑细胞中NPY主要在细胞质中表达;不同部位脑组织中主要以前脑的视前区、视顶盖和脑垂体,小脑的豆状核区域以及延脑的尾状核区域表达量较丰富。进一步进行饥饿补偿验证发现,饥饿可引起脑组织NPY表达量显著升高,摄食后NPY表达量显著下降。上述结果表明,NPY基因可能在兰州鲇生长和摄食过程中都具有重要作用,可作为对后续功能基因进行编辑的重要验证并创制新种质,对加快兰州鲇良种新品种选育具有重要的理论意义和技术支撑作用。 展开更多
关键词 兰州鲇 NPY基因 生物信息学 基因克隆 原位杂交 组织表达
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贵州杂交黄牛MSTN基因cDNA克隆、生物信息学及多态性分析 被引量:1
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作者 王可人 游绍航 +1 位作者 主性 田兴贵 《家畜生态学报》 北大核心 2023年第4期13-21,共9页
为比较贵州杂交黄牛肌肉生长抑制素(Myostatin,MSTN)性质、结构和多态性与本地黄牛的差别,该研究先通过RT-PCR克隆MSTN基因cDNA并进行生物信息学分析,再通过PCR-SSCP结合直接测序法筛查贵州本地黄牛和杂交黄牛MSTN基因外显子多态性。结... 为比较贵州杂交黄牛肌肉生长抑制素(Myostatin,MSTN)性质、结构和多态性与本地黄牛的差别,该研究先通过RT-PCR克隆MSTN基因cDNA并进行生物信息学分析,再通过PCR-SSCP结合直接测序法筛查贵州本地黄牛和杂交黄牛MSTN基因外显子多态性。结果在贵州杂交黄牛MSTN基因CDS区序列发现4个突变位点,其中3个为错义突变:76 bp处G/A替换,引起天冬酰胺变为天冬氨酸;79 bp处G/A替换,引起丝氨酸变为甘氨酸;1066 bp处G/A替换,引起谷氨酸变为赖氨酸。267 bp处G/A替换与525 bp处A/G替换,为同义突变。MSTN蛋白基本理化性质和结构与普通牛基本一致,但有细微的差别;通过基因同源性和系统进化树,结合motif、蛋白保守域和结构域分析,可知MSTN基因在哺乳动物中高度保守,但也存在遗传多样性。贵州本地黄牛外显子发现3个SNP位点,分别为g.6279107 G>C、g.6281238 T>C和g.6283933 A>C,贵州杂交黄牛中未发现多态位点。单倍型分析共发现6种单倍型和14种双倍型,双倍型H3H5频率最高,且杂交牛的双倍型均为H3H5,之后可一步将此与生长性状进行关联分析研究。 展开更多
关键词 贵州杂交黄牛 肌肉生长抑制素基因 基因克隆 生物信息学 多态位点
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大豆Glyma04G227700生物信息学分析及与Glyma08G11030互作研究 被引量:1
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作者 罗飞 刘灿 +1 位作者 于月华 倪志勇 《大豆科学》 CAS CSCD 北大核心 2023年第2期182-187,共6页
咖啡酸-O-甲基转移酶(caffeic acid-O-methyltransferase, COMT)是一种催化苯丙素类化合物羟基上氧原子的甲基化酶。前期研究表明,大豆抵御干旱胁迫的F-box基因Glyma08G11030编码蛋白可能与COMT蛋白Glyma04G227700发生互作,为了预测分... 咖啡酸-O-甲基转移酶(caffeic acid-O-methyltransferase, COMT)是一种催化苯丙素类化合物羟基上氧原子的甲基化酶。前期研究表明,大豆抵御干旱胁迫的F-box基因Glyma08G11030编码蛋白可能与COMT蛋白Glyma04G227700发生互作,为了预测分析大豆COMT基因Glyma04G227700的功能及二者的互作关系,本研究克隆Glyma04G227700基因,并对其进行生物信息学分析,通过实时荧光定量检测方法分析其在大豆不同组织中的表达情况,并利用酵母双杂交系统验证蛋白互作情况。结果显示:Glyma04G227700基因全长1 095 bp,编码366个氨基酸,与野大豆(RZC17879.1 Glycine soja)亲缘关系较近。Glyma04G227700在大豆的根、茎、叶、子叶中都有表达,且在根和茎中表达量较高,在子叶中表达量最低。酵母双杂结果表明Glyma08G11030与Glyma04G227700蛋白不存在互作。 展开更多
关键词 大豆 咖啡酸-O-甲基转移酶(COMT) Glyma04G227700 基因克隆 生物信息学分析 酵母双杂 Glyma08G11030
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黄鳝细胞质动力蛋白3基因的克隆及其在性逆转中的功能分析
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作者 连子童 夏雪平 +3 位作者 李忠 田海峰 蒋钦杨 胡乔木 《淡水渔业》 CSCD 北大核心 2023年第3期93-102,共10页
为解析黄鳝(Monopterus albus)性逆转机制,实验以雌、雄、间性发育黄鳝为研究对象,分析不同组织、不同发育时期性腺、甲基睾丸酮处理性腺及Zebularine处理性腺原代细胞后dynlt3基因表达模式的变化及其在性腺中的表达定位。性腺转录组测... 为解析黄鳝(Monopterus albus)性逆转机制,实验以雌、雄、间性发育黄鳝为研究对象,分析不同组织、不同发育时期性腺、甲基睾丸酮处理性腺及Zebularine处理性腺原代细胞后dynlt3基因表达模式的变化及其在性腺中的表达定位。性腺转录组测序结果显示,基因全长1082 bp,开放阅读框354 bp,编码117个氨基酸。生物信息学分析显示,dynlt3基因编码蛋白质的二级结构包含37.96%的α-螺旋,29.20%的β-折叠,32.85%的无规则卷曲。系统进化分析结果显示,黄鳝DYNLT3氨基酸序列与硬骨鱼纲中底鳉(Fundulus heteroclitus)同源性最高。实时荧光定量PCR结果表明,dynlt3基因在黄鳝肌肉和脑具有较高表达,心脏次之,在其它各组织表达量较低。在性腺的不同发育时期,在间性后期和雄性中表达量显著性高于雌性与间性早期。甲基睾酮处理后黄鳝卵巢组织结构发生明显退化,卵母细胞退化且数量减少,结缔组织间出现空泡结构;dynlt3基因在卵巢中表达量显著性下调。原位杂交分析dynlt3基因在性腺组织中的表达定位结果显示,在不同性腺发育时期均检测到dynlt3阳性信号,间性性腺组织中dynlt3基因主要在精原细胞和初级精母细胞中表达,精巢组织中dynlt3基因可在精原细胞与初级精母细胞中,卵巢组织阳性信号较弱,主要在II时相卵母细胞细胞质中表达。Zebularine处理性腺原代细胞后,dynlt3基因表达无显著性差异。以上研究表明,dynlt3参与黄鳝性腺发育过程,并在黄鳝性逆转及性腺细胞发育过程中发挥重要作用,但是甲基化可能不参与其基因表达调控。 展开更多
关键词 黄鳝(Monopterus albus) 动力蛋白轻链3(dynlt3) 克隆 原位杂交 表达
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池蝶蚌HsPif基因的克隆及表达分析
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作者 熊雅萍 马慧妹 +2 位作者 胡蓓娟 邱齐骏 洪一江 《水生生物学报》 CAS CSCD 北大核心 2023年第2期339-344,共6页
为了探究池蝶蚌(Hyriopsis schlegelii)酸性基质蛋白Pif基因的基本功能,研究通过RACE-PCR技术首次在池蝶蚌中获得了Pif基因的cDNA全长序列,命名为HsPif,其全长为3457 bp, 5′端非翻译区(5′UTR)为485 bp,3′端非翻译区(3′UTR)为363 bp... 为了探究池蝶蚌(Hyriopsis schlegelii)酸性基质蛋白Pif基因的基本功能,研究通过RACE-PCR技术首次在池蝶蚌中获得了Pif基因的cDNA全长序列,命名为HsPif,其全长为3457 bp, 5′端非翻译区(5′UTR)为485 bp,3′端非翻译区(3′UTR)为363 bp,开放阅读框(ORF)3072 bp,共编码1023个氨基酸,生物信息学分析结果显示HsPif蛋白含有一个von-Willebrand因子A型结构域和三个几丁质结合结构域;氨基酸组成成分结果表明天冬氨酸含量最高,组氨酸含量最低;HsPif蛋白亲水指数为–0.566,为亲水蛋白。构建系统进化树分析显示Pif基因保守性较高,与三角帆蚌(Hyriopsis cumingii)Pif相似度最高,且与其他贝类在同一个大支上。组织荧光定量PCR结果表明:HsPif基因主要在外套膜中表达;分子原位杂交显示杂交反应主要在外套膜上皮细胞发生。进行植核手术后进行qPCR检测HsPif基因的表达量变化,实验结果表明, HsPif基因可能与珍珠层的分泌有关,有助于进一步了解珍珠形成的机制,为珍珠养殖提供参考。 展开更多
关键词 HsPif 基因克隆 原位杂交 植核 组织表达 珍珠囊 池蝶蚌
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尾叶桉×细叶桉杂种无性系扦插生根和生长性状的研究 被引量:14
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作者 甘四明 李梅 +4 位作者 李发根 吴坤明 吴菊英 卢国桓 白嘉雨 《林业科学研究》 CSCD 北大核心 2006年第2期135-140,共6页
对尾叶桉×细叶桉5个杂种组合的181个无性系扦插生根和生长性状的研究表明:每穗根数、每穗最长根长和生根率3个扦插生根性状,杂种组合间和杂种组合内无性系间的差异均达0.01显著水平;46个月生树高、胸径和单株材积3个生长性状... 对尾叶桉×细叶桉5个杂种组合的181个无性系扦插生根和生长性状的研究表明:每穗根数、每穗最长根长和生根率3个扦插生根性状,杂种组合间和杂种组合内无性系间的差异均达0.01显著水平;46个月生树高、胸径和单株材积3个生长性状,杂种组合问差异不显著,杂种组合内无性系间的差异达0.01显著水平。无性系扦插生根性状和生长性状的广义遗传力介于0.32(每穗最长根长)~0.94(生根率)之间,表明各性状受中等到高等程度的遗传控制。无性系扦插生根性状平均值与生长性状平均值的表型相关均不显著。无性系各生长性状在13、18、46个月生之间的年年相关均达到0.01显著水平,表明在较早生长期进行无性系选择具有一定的可行性。本研究初选出易扦插、速生的尾细桉杂种无性系24个,无性系间单株材积株间变异系数为13.87%~68.55%,无性系内变异可能与母株年龄效应、位置效应和立地环境条件等C-效应因子有关。对桉树杂交育种而言,杂交制种后直接进行育苗、扦插和无性系试验是一条快捷有效的无性系培育途径。 展开更多
关键词 桉树 杂种 无性系 扦插 生长 广义遗传力
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白杨派杂种无性系植株早期性状变异与选择研究 被引量:26
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作者 赵曦阳 马开峰 +4 位作者 沈应柏 张明 李奎友 邬荣领 张志毅 《北京林业大学学报》 CAS CSCD 北大核心 2012年第2期45-51,共7页
以30个白杨杂种无性系为材料,测定树高、胸径、树形、叶片、树皮特性等17个指标。方差分析与遗传参数计算结果表明,白杨杂种无性系各指标差异极显著(P<0.001),各指标表型变异系数变化范围为15.63%~57.50%,遗传变异系数变化范围为8.... 以30个白杨杂种无性系为材料,测定树高、胸径、树形、叶片、树皮特性等17个指标。方差分析与遗传参数计算结果表明,白杨杂种无性系各指标差异极显著(P<0.001),各指标表型变异系数变化范围为15.63%~57.50%,遗传变异系数变化范围为8.99%~52.57%,重复力高(0.7734~0.9848)。说明白杨杂种无性系各性状存在广泛变异,且这种变异受较强的遗传因素控制。表型相关结果表明,白杨杂种无性系树高、地径、胸径、单株材积和冠幅之间均达显著相关水平,遗传相关系数接近表型相关系数。主成分分析结果显示,白杨杂种无性系14个性状前3个主成分的累积贡献率为84.93%,其中第1主成分为生长量等性状,第2主成分为干形等性状,第3主成分为叶片性状。利用综合评定法对无性系进行评价,当入选率为10%时,无性系BL204、BL206和BL207入选,入选无性系树高、地径、胸径和单株材积的遗传增益分别为27.41%、31.73%、32.90%和102.37%。这些分析结果为下一步选育优良无性系提供了重要的信息和依据。 展开更多
关键词 白杨 杂种无性系 遗传变异
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