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991.
S-Adenosyl-L-methionine: uroporphyrinogen III methyltransferase (SUMT), a key regulatory enzyme, converts uroporphyrinogen III to precorrin-2 in the porphinoids biosynthesis. In this study, the mature SUMT was signified that the maize SUMT precursor encoded by the open reading frame of maize SUMT cDNA was deleted the first 91 amino acids constituting the postulated signal peptide. Several mature SUMT fusion and deletion mutants were conducted. It actively expressed in Escherichia coli that the mature SUMT, or the truncated one deleting the C-terminal extra 52 amino acids based on SUMT sequence comparisons. On the contrary, it expressed as an inclusion body in E. coli that the mature SUMT fusion mutant, the SUMT precursor, or the mature SUMT deleting the N-terminal 36 amino acids including glycine-rich region involved directly in SAM binding. The purified His6-tagged mature SUMT was homodimer with a molecular weight of 34 kDa, as shown by SDS-PAGE, 52 kDa using gel-filtration chromatography, and 79 kDa by dynamic light scattering assay. Red fluorescent compounds were associated with the recombinant mature SUMT which were identified as sirohydrochlorin and trimethylpyrrocorphin by spectroscopic analysis. This association slightly altered the protein secondary structure confirmed by circular dichroism assay. 相似文献
992.
Li X Wang Q Zheng Y Lv S Ning S Sun J Huang T Zheng Q Ren H Xu J Wang X Li Y 《Nucleic acids research》2011,39(22):e153
The identification of human cancer-related microRNAs (miRNAs) is important for cancer biology research. Although several identification methods have achieved remarkable success, they have overlooked the functional information associated with miRNAs. We present a computational framework that can be used to prioritize human cancer miRNAs by measuring the association between cancer and miRNAs based on the functional consistency score (FCS) of the miRNA target genes and the cancer-related genes. This approach proved successful in identifying the validated cancer miRNAs for 11 common human cancers with area under ROC curve (AUC) ranging from 71.15% to 96.36%. The FCS method had a significant advantage over miRNA differential expression analysis when identifying cancer-related miRNAs with a fine regulatory mechanism, such as miR-27a in colorectal cancer. Furthermore, a case study examining thyroid cancer showed that the FCS method can uncover novel cancer-related miRNAs such as miR-27a/b, which were showed significantly upregulated in thyroid cancer samples by qRT-PCR analysis. Our method can be used on a web-based server, CMP (cancer miRNA prioritization) and is freely accessible at http://bioinfo.hrbmu.edu.cn/CMP. This time- and cost-effective computational framework can be a valuable complement to experimental studies and can assist with future studies of miRNA involvement in the pathogenesis of cancers. 相似文献
993.
Yu‐Kuang Liao Maël Brossard Dan‐Hua Hsieh Tzu‐Neng Lin Martin D. B. Charlton Shun‐Jen Cheng Chyong‐Hua Chen Ji‐Lin Shen Lung‐Teng Cheng Tung‐Po Hsieh Fang‐I Lai Shou‐Yi Kuo Hao‐Chung Kuo Pavlos G. Savvidis Pavlos G. Lagoudakis 《Liver Transplantation》2015,5(2)
A novel scheme for hybridizing inkjet‐printed thin film Cu(In,Ga)Se2 (CIGS) solar cells with self‐assembled clusters of nanocrystal quantum dots (NQDs), which provides a 10.9% relative enhancement of the photon conversion efficiency (PCE), is demonstrated. A non‐uniform layer of NQD aggregates is deposited between the transparent conductive oxide and a CdS/CIGS p‐n junction using low cost pulsed‐spray deposition. Hybridization significantly improves the external quantum efficiency of the hybrid devices in the absorption range of the NQDs and in the red to near‐IR parts of the spectrum. The low wavelength response enhancement is found to be induced by luminescent down‐shifting (LDS) from the NQD layer, while the increase at longer wavelengths is attributed to internal scattering from NQD aggregates. LDS is demonstrated using time‐resolved spectroscopy, and the morphology of the NQD layer is investigated in fluorescence microscopy and cross‐sectional transmission electron microscopy. The influence of the NQD dose on the PCE of the hybrid devices is investigated and an optimum value is obtained. The low costs and limited material consumptions associated with pulsed‐spray deposition make these flexible hybrid devices promising candidates to help push thin‐film photovoltaic technology towards grid parity. 相似文献
994.
顽拗植物龙眼基因组DNA提取方法的研究 总被引:20,自引:2,他引:20
为从顽拗植物龙眼(Dimocarpus longan Lour.)叶片中获得可供后续分子生物学操作的基因组DNA.针对其组织细胞内富含多酚、多糖、单宁及色素等物质的特点,采用改进的CTAB法和SDS法,即在核裂解之前先破碎细胞.将存在于细胞质中的次生物质去除后再裂解细胞桉.结合其它一些改进措施.提取到的DNA沉淀呈纯白色.极易溶解于TE中。两种改进方法的OD260和OD280比值分别达到1.82和1.73,其鲜叶基因组DNA产量分别为103ug/g和127ug/g:RAPD扩增条带清晰,丰富.完全满足后续分子生物学操作的要求,其中改良CTAB方法效果更为理想,与之埘比.传统的CTAB法和SDS法提取到的DNA沉淀呈浅黄色甚至红褐色,很难被TE溶解,其OD260和OD280比值均低于1.5,也得不到扩增产物。 相似文献
995.
土壤中脂肪酶产生菌的筛选、鉴定及脂肪酶基因的克隆 总被引:2,自引:0,他引:2
以橄榄油为惟一碳源进行富集培养、以罗丹明B为指示剂的平板进行初筛,摇瓶复筛得到产脂肪酶菌株;利用滴定法、平板扩散法以及转酯反应试验,最终筛选出具有较高转酯活性脂肪酶的菌株B2.通过对B2进行形态学观察,生理生化测定以及16S rDNA特征片段比较分析,初步确定B2为克雷伯氏菌属(Klebsiella sp.).通过特定引物PCR扩增得到B2菌株两个脂肪酶基因K1和K2.利用丙酮法和(NH)2SO4沉降法得到B2菌株体内的胞内酶,以叔丁醇为溶剂,催化棉籽油与甲醇反应,经过薄层层析和高效液相检测产物为生物柴油. 相似文献
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997.
原生质体的大量制备是研究原生质体转化、诱变、融合等技术的关键,液泡的制备对研究液泡中分解、转运有机物的特性具有重要意义,但目前分离技术还不够成熟.本研究从聚多曲霉菌菌丝中分离原生质体和液泡并对分离条件进行优化,以聚多曲霉菌DJ515-2菌丝为材料,探索不同因素对原生质体和液泡制备分离效果的影响.结果表明,聚多曲霉菌DJ515-2在菌龄42 h,以3%纤维素酶、1%蜗牛酶和3%的溶壁酶组成复合酶液,25℃下酶解4 h,原生质体达到最大产量,为5.167×105个/mL.同时,在此基础上裂解液泡的最优条件为pH7.5、1.0mol/L KCl、0.020%Triton X-10,其产量达2.63×105个/mL,产率为原生质体的50.8%,相比采用多元碱化合物诱导真菌原生质体裂解释放液泡的产率增加了 40%~45%.本研究为真菌和植物的各种原生质体技术及基于亚细胞层面的研究提供了材料基础. 相似文献
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