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61.
62.
Pyrroloquinoline quinone (PQQ) is a versatile quinone cofactor participating in numerous biological processes. Klebsiella pneumoniae can naturally synthesize PQQ for harboring intact PQQ synthesis genes. Previous metabolic engineering of K. pneumoniae failed to overproduce PQQ due to the employment of strong promoter in expression vector. Here we report that a moderate rather than strong promoter is efficient for PQQ production. To screen an appropriate promoter, a total of four distinct promoters—lac promoter, pk promoter of glycerol dehydratase gene (dhaB1), promoter of kanamycin resistance gene, and T7 promoter (as the control)—were individually used for overexpressing the endogenous PQQ genes in K. pneumoniae along with heterologous expression in Escherichia coli. We found that all recombinant K. pneumoniae strains produced more PQQ than recombinant E. coli strains that carried corresponding vectors, indicating that K. pneumoniae is superior to E. coli for the production of PQQ. Particularly, the recombinant K. pneumoniae recruiting the promoter of kanamycin resistance gene produced the highest PQQ (1,700 nmol), revealing that a moderate rather than strong promoter is efficient for PQQ production. Furthermore, PQQ production was roughly proportional to glucose concentration increasing from 0.5 to 1.5 g/L, implying the synergism between PQQ biosynthesis and glucose utilization. This study not only provides a feasible strategy for production of PQQ in K. pneumoniae, but also reveals the exquisite synchronization among PQQ biosynthesis, glucose metabolism, and cell proliferation. 相似文献
63.
Century-Scale Responses of Ecosystem Carbon Storage and Flux to Multiple Environmental Changes in the Southern United States 总被引:2,自引:0,他引:2
Hanqin Tian Guangsheng Chen Chi Zhang Mingliang Liu Ge Sun Arthur Chappelka Wei Ren Xiaofeng Xu Chaoqun Lu Shufen Pan Hua Chen Dafeng Hui Steven McNulty Graeme Lockaby Eric Vance 《Ecosystems》2012,15(4):674-694
Terrestrial ecosystems in the southern United States (SUS) have experienced a complex set of changes in climate, atmospheric CO2 concentration, tropospheric ozone (O3), nitrogen (N) deposition, and land-use and land-cover change (LULCC) during the past century. Although each of these factors has received attention for its alterations on ecosystem carbon (C) dynamics, their combined effects and relative contributions are still not well understood. By using the Dynamic Land Ecosystem Model (DLEM) in combination with spatially explicit, long-term historical data series on multiple environmental factors, we examined the century-scale responses of ecosystem C storage and flux to multiple environmental changes in the SUS. The results indicated that multiple environmental changes shifted SUS ecosystems from a C source of 1.20?±?0.56?Pg (1?Pg?=?1015?g) during the period 1895 to 1950, to a C sink of 2.00?±?0.94?Pg during the period 1951 to 2007. Over the entire period spanning 1895–2007, SUS ecosystems were a net C sink of 0.80?±?0.38?Pg. The C sink was primarily due to an increase in the vegetation C pool, whereas the soil C pool decreased during the study period. The spatiotemporal changes of C storage were caused by changes in multiple environmental factors. Among the five factors examined (climate, LULCC, N deposition, atmospheric CO2, and tropospheric O3), elevated atmospheric CO2 concentration was the largest contributor to C sequestration, followed by N deposition. LULCC, climate, and tropospheric O3 concentration contributed to C losses during the study period. The SUS ecosystem C sink was largely the result of interactive effects among multiple environmental factors, particularly atmospheric N input and atmospheric CO2. 相似文献
64.
Ge X McFarlane C Vajjala A Lokireddy S Ng ZH Tan CK Tan NS Wahli W Sharma M Kambadur R 《Cell research》2011,21(11):1591-1604
TGF-β and myostatin are the two most important regulators of muscle growth. Both growth factors have been shown to signal through a Smad3-dependent pathway. However to date, the role of Smad3 in muscle growth and differentiation is not investigated. Here, we demonstrate that Smad3-null mice have decreased muscle mass and pronounced skeletal muscle atrophy. Consistent with this, we also find increased protein ubiquitination and elevated levels of the ubiquitin E3 ligase MuRF1 in muscle tissue isolated from Smad3-null mice. Loss of Smad3 also led to defective satellite cell (SC) functionality. Smad3-null SCs showed reduced propensity for self-renewal, which may lead to a progressive loss of SC number. Indeed, decreased SC number was observed in skeletal muscle from Smad3-null mice showing signs of severe muscle wasting. Further in vitro analysis of primary myoblast cultures identified that Smad3-null myoblasts exhibit impaired proliferation, differentiation and fusion, resulting in the formation of atrophied myotubes. A search for the molecular mechanism revealed that loss of Smad3 results in increased myostatin expression in Smad3-null muscle and myoblasts. Given that myostatin is a negative regulator, we hypothesize that increased myostatin levels are responsible for the atrophic phenotype in Smad3-null mice. Consistent with this theory, inactivation of myostatin in Smad3-null mice rescues the muscle atrophy phenotype. 相似文献
65.
泡沙参复合体(桔梗科)的物种生物学研究——Ⅲ.性状的遗传变异及其分类价值 总被引:5,自引:0,他引:5
阐明性状变异的遗传基础是评估性状分类价值以及揭示类群关系和进化过程的前提。本文根据泡沙参复合体内6个天然居群的材料,利用居群样本分析、子代测定和杂交试验等手段,结合统计学方法对该复合体一些在以往类群划分中起鉴别作用的形态性状进行了遗传学分析。结果表明,叶形、叶缘锯齿数目和大小、茎叶被毛、花萼裂片齿数、花梗长短等性状都是遗传性比较强的性状,而且性状之间有很强的相关性。但是,这些性状在居群内呈现连续变异且变异幅度非常大,以致于变异的两个极端个体被分别作为不同的种处理。通过对野外居群样本的统计分析和对极端变异类型个体所进行的子代测定及其人工杂交,进一步证实上述性状是受多基因决定的数量性状,其变异幅度大,变异式样中并不存在任何程度的间断性,因而至少在本复合体内没有鉴别意义。因此,以往建立在上述性状变异基础上的3个类群(A.biformifolia Y.Z.Zhao;A.bockiana Diels和A.polydentata P.F.Tu et G.I.Xu)均不成立。最后,本文对形态性状变异的遗传基础及其与类群划分的关系进行了讨论。 相似文献
66.
The inhibition of lung cancer cell growth by intracellular immunization with LC—1 ScFv 总被引:2,自引:0,他引:2
INTRODUCTIONLung cancer remains the leading cause of can-cer mortaIity in the world, accounting for more thanone sixth of cancer deaths in the world[1]. Antibod-ies have been proved to be a powerful tool fOr thestudy of 1ung cancer. A monoclonal IgM antibody,LC-1, was obtained in our laboratory. It can reactat a high rate with all four pathological types of lungcancers, including lung adenocarcinoma, 1ung squamous carcinoma, large cell lung cancer and smaIlcell lung cancert but not wit… 相似文献
67.
表达流感病毒神经氨酸酶基因的重组腺病毒的构建 总被引:1,自引:0,他引:1
通过RT-PCR方法扩增流感病毒神经氨酸酶基因,将其克隆到腺病毒穿梭载体pTrackCMV,此重组质粒与腺病毒DNA共转化大肠杆菌BJ5183,通过细菌内同源重组获得重组腺病毒DNA,将其转染293细胞获得重组腺病毒.经PCR证实目的基因已整合至腺病毒基因组中,western blot检测到神经氨酸酶的表达.重组病毒经滴鼻和灌胃两种途径免疫小鼠,结果表明2次免疫后滴鼻组和灌胃组均产生明显的免疫应答反应,滴鼻组的免疫效果优于灌胃组. 相似文献
68.
目的探讨慢性胃炎胃粘膜肠化生CD44、CD44V6及cyclin D1、Cyclin E表达的意义。方法利用免疫细胞化学技术对39例伴有肠化生的慢性胃炎和5例正常人胃窦粘膜的活检组织进行检查。结果正常人胃窦粘膜上皮,腺上皮对CD44、CD44V6、Cyclin D1和Cyclin E均为阴性,但在有神经内分泌样细胞的粘膜,CD44V6和cyclin D1为阳性。慢性胃炎肠化生区和不典型增生区除CD44为阴性外,CD44V6、cyclin D1和cyclin E均呈现不同的阳性反应,但未见有阳性的神经内分泌样细胞。间质细胞大都呈阳性反应。结论CD44V6、cyclin D1和cyclin E可能是胃癌前状态的早期事件,而CD44可能为胃癌晚期的标志物。 相似文献
69.
隐甲藻深层培养产生二十碳五烯酸的研究 总被引:5,自引:0,他引:5
二十碳五烯酸 (C2 0 :5 ,Eicosapentaenoicacid ,简称EPA)是一种重要的ω 3系列人体必需多不饱和脂肪酸 (Polyun saturatedfattyacids,PUFA) [1~ 2 ] 。自从Dyerberge等人报道了二十碳五烯酸对防止和治疗血栓、关节硬化、抗炎症、抗癌、促进脑组织发育等方面具有明显效果以来 ,人们对它的营养和医药价值及生产方法进行了广泛的研究[3~ 6] 。目前 ,二十碳五烯酸主要来源于深海鱼油 ,但其产量不稳定 ,纯化工艺复杂且含有难以消除的鱼腥味 ,难以满足市场的需求。由于二十碳五… 相似文献
70.
Guan S Ma S Zhu Y Ge R Wang Q Wang JH 《Biochemical and biophysical research communications》2006,345(1):175-180
Cerebellum is involved in the motion coordination and working memory, to which the programming of sequential spikes at Purkinje cells is essential. It is not clear about the intrinsic mechanisms underlying spike capacity and timing precision as well as their postnatal maturation. We investigated the programming and intrinsic property of sequential spikes at Purkinje neurons during postnatal development by whole-cell recording in cerebellar slices. Cerebellar Purkinje neurons demonstrate the increasing of spike capacity and timing precision, as well as the lowering of refractory periods and threshold potentials during the postnatal maturation. In addition, the correlation between spike parameters and intrinsic properties converts to be more linear. This postnatal plasticity of neuronal intrinsic properties improves the timing precision and capacity of spike programming at cerebellar Purkinje neurons. 相似文献