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    题名 作者 年代 出处 被引量
1过量表达沙冬青巯基蛋白酶抑制剂基因AmPI提高大肠杆菌低温与热胁迫抗性显示文摘沙冬青是我国北方内蒙古荒漠地区特有的常绿阔叶灌木,具有很强的抗低温、干旱、盐碱能力,是重要的抗逆基因资源植物.将从低温诱导的沙冬青幼苗中克隆获得的巯基蛋白酶抑制剂基因AmPI亚克隆后构建到原核表达载体pTWIN1上,得到重组表达载体pT-PI,双酶切及测序结果证明pT-PI已经构建成功.将pT-PI转入大肠杆菌ER2566中,经异丙基β-硫代半乳糖苷(IPTG)诱导后,SDS-PAGE电泳检测发现有Mr35×103的目的融合蛋白表达,与预测结果一致,其中以在37℃诱导3~4h条件下表达量最高,表明目的基因AmPI在大肠杆菌中得到了成功表达.分别测定了低温(0℃)和热胁迫(50℃)条件下AmPI宿主菌的生存能力,结果表明AmPI的表达能提高宿主菌的存活率,可能是AmPI对细胞具有保护功能.刘瑞玲 刘美芹 史军娜 陈玉珍 卢存福 2010应用与环境生物学报2010,16,3:18
2Variation of vertical and horizontal drilling rates depending on some rock properties in the marble quarries显示文摘The main objective of this study is to determine the rates of vertically and horizontally oriented drilling processes in marble quarries and to observe the factors affecting the drilling rates in terms of physical and mechanical properties of the rocks. In situ drilling tests were performed in different marble quarries with different marble types and drilling times and penetration rates for a series of successive depthincrements were trying to be determined under vertically and horizontally oriented conditions. In order to understand the relation between the parameters that are investigated within the scope of this research, uniaxial compressive strength, Brazilian tensile strength, impact strength, Bohme abrasion strength, P-wave velocity, porosity, unit volume weight, Schmidt hardness index and brittleness index values were correlated with the drilling rates. It was noticed that the porosity and unit volume weight could be taken as the key parameters among them for obtaining meaningful correlation with drilling performance. It was also observed that the physical and mechanical rock properties are more relevant in vertical drilling than horizontal drilling.Servet Demirdag Nazmi Sengun Ibrahim Ugur Tamer Efe Deniz Akbay Rasit Altindag 2014International Journal of Mining Science and Technology2014,24,2:7
3Bowman-Brik胰蛋白酶抑制剂在水稻叶片生长和种子萌发过程中的表达显示文摘Bowman-Birk胰蛋白酶抑制剂(BBTI)是一类富含半胱氨酸的丝氨酸蛋白酶抑制剂,广泛分布于豆科、禾本科和菊科植物中,在蛋白质存储和植物防御中具有重要作用。水稻(Oryza sativa L.ssp.indica)BBTI家族具有13个成员,其中BBTI-9和BBTI-10都为假基因。其余BBTI成员都存在数量不同的保守Bowman-Birk结构域。为了解BBTI的表达信息,本研究主要通过设计多肽序列的方法,制备了11个BBTI的多克隆抗体,利用Western blot技术检测其在叶片生长发育和种子萌发过程中的表达。结果表明BBTI在叶片生长过程中呈现不同的时空特异性表达,BBTI-1、-2、-11、-12和-13伴随叶片生长含量增加,BBTI-5、-6和-8在苗期表达含量高,而BBTI-3、-4、-7在苗早期1cm时表达量达到峰值,以修饰或多聚体的形式在叶片生长过程中表达。在种子萌发过程中发现BBTI-8在种子中表达量较高,萌发后2h降到最低,24h后一条分子量略小的降解带开始表达,其余抗体在种子萌发阶段未发现有明显的变化。本研究还统计了苗期叶片、分蘖期叶片和萌发期的种子的MPSStags数据,试图比较转录与蛋白质水平对应关系。本研究结果证明,BBTI蛋白质在水稻生长发育过程中存在差异表达,为深入了解其功能提供了重要线索。徐文静 李莉云 史佳楠 李静宜 巩校东 刘丽娟 刘国振 2012农业生物技术学报2012,20,3:4
4Cloning of Proteinase Inhibitor Gene StPI in Diploid Potato and Its Expression Analysis显示文摘A full-length cDNA of proteinase inhibitor gene with completed open reading frame of 116 amino acids was cloned from Ralstonia solanacearum (Rs) resistant potato leaves using the rapid amplification of cDNA ends (RACE) method and designated as StPI. BLAST search against NCBI showed that the StPI gene shared 89% identity with potato proteinase inhibitor I precursor in nucleotide and 74% in amino acid. Analysis of semi-quantitative RT-PCR indicated that this gene was induced by Rs as well as up-regulated by jasmonic acid (JA). The StPI gene expression reached the highest level during 6-12 h post Rs-inoculation or JA-treatment,and then leveled off. Moreover,this gene was strongly induced by JA and its mRNA accumulation increased more quickly than that of Rs-inoculation. The StPI gene may play a role in potato resistance against Rs. The induction of StPI by Rs invasion may have a similar signal transduction pathway with JA treatment.LI Guang-cun JINLi-ping XIE Kai-yun LI Ying QU Dong-yu 2007Agricultural Sciences in China2007,6,11:3
5Molecular characterization and developmental expression of the gene encoding the prothoracicotropic hormone in the beet armyworm, Spodoptera exigua显示文摘Prothoracicotropic hormone (PTTH), a neuropeptide hormone stimulating the prothoracic glands to synthesize ecdysone, plays an important role in regulating postembryonic development in insects. The cDNA encoding PTTH was isolated and sequenced from the beet armyworm, Spodoptera exigua (Spe). The deduced a?mino acid sequence is composed of a signal peptide, a peptide (65 amino acids) of un-known function, and a mature PTTH molecule (111 amino acids). The Spe-PTTH shows similarities (45.5%―70.3%) to other known PTTHs reported in Lepidoptera species, but 7 cysteine r?esidues and the hydrophobic regions were conserved. Whole-mount immunocytochemistry by using an antiserum against recombinant Helicoverpa armigera PTTH showed that Spe-PTTH was synthesized in two pairs of neurosecretory cells in the S. exigua brain. Northern blot analysis demonstrates the presence of a 1.2-kb transcript in the brain. The Spe-PTTH mRNA is detectable at high levels at the wandering larval stage, early pupal stage, and pharate adult stage, suggesting that the Spe-PTTH gene might be corre-lated with molting, metamorphosis, and reproduction.XU Jun1,SU JianYa3, SHEN JinLiang3 & XU WeiHua1,2 1 Department of Molecular and Cell Biology, School of Life Sciences, University of Science and Technology of China, Hefei 230027, China 2 State Key Laboratory of Biocontrol and Institute of Entomology, School of Life Sciences, Zhongshan University, Guangzhou 510275, China 3 Department of Plant Protection, Nanjing Agriculture University, Nanjing 210095, China 2007Science China(Life Sciences)2007,50,4:3
6马铃薯蛋白酶抑制子StPI基因的克隆及表达分析显示文摘【目的】克隆马铃薯蛋白酶抑制子基因StPI的全长cDNA。【方法】以马铃薯高抗青枯病二倍体基因型ED13为材料,采用RACE方法进行StPI基因全长cDNA的克隆,利用半定量RT-PCR方法进行该基因的诱导表达分析。【结果】获得了StPI基因的全长cDNA,序列分析表明:该基因具有完整的开放阅读框架,编码116个氨基酸,与马铃薯蛋白酶抑制子I前体具有较高的同源性(核苷酸和氨基酸序列同源性分别为89%和74%)。该基因同时受青枯病菌的诱导和茉莉酸的调节,在6~12h内即达到最高表达水平,但二者又有明显不同。相对而言,StPI基因受青枯病菌的诱导较弱,而受茉莉酸(JA)的诱导较为强烈,在JA处理3h表达量即明显升高,6~12h迅速升至最高。【结论】本研究从马铃薯抗青枯病基因型ED13中获得了StPI基因的全长cDNA。该基因可能参与了马铃薯的抗青枯病反应,且病菌处理对该基因的诱导可能与JA刺激具有相似或相同的信号途径。李广存 金黎平 谢开云 李颖 屈冬玉 2007中国农业科学2007,40,9:2
7小麦一个新BBI型蛋白酶抑制剂基因TaUES的克隆及初步分析显示文摘根据小麦基因表达芯片结果,以山融3号小麦叶片为试验材料,克隆获得在盐胁迫处理24 h时表达显著提高的基因TaUES(up-regulated expression under saline-stress in wheat,ID:KC408382)。序列分析显示,TaUES编码一个富含半胱氨酸的97个氨基酸的多肽,该多肽含有1个BowB结构域和10个保守的半胱氨酸残基;与小麦或其他物种BBI型蛋白酶抑制剂氨基酸序列具有较高的同源性。推测TaUES是小麦中一种新的BBI型蛋白酶抑制剂基因。基因表达分析表明,TaUES基因参与盐和干旱胁迫应答。异源过表达转基因功能初步分析表明,过表达TaUES转基因烟草后代株系耐盐性提高。徐平丽 彭振英 马小凤 唐桂英 单雷 夏光敏 2014植物遗传资源学报2014,15,2:2
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