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[目的]研究甲基化修饰对遗传基因调节的作用。[方法]利用拟南芥RNA反转录得到的cDNA序列为模板,克隆出SUVH6-SRA基因,并成功构建了含有SUVH6-SRA基因的表达载体pMAL-c2P-SUVH6-SRA,然后将pMAL-c2P-SUVH6-SRA质粒转化到大肠杆菌BL21（DE3）中,诱导表达带有MBP标签的SUVH6-SRA融和蛋白。[结果]pMAL-c2P-SUVH6-SRA克隆在转化到大肠杆菌BL21（DE3）后,在37℃条件下,经过终浓度为1.0 mmol/L的IPTG诱导,大肠杆菌BL21（DE3）能够获得大量可溶性MBP-SUVH6-SRA融和蛋白并顺利表达,SDS-PAGE显示诱导表达的融和蛋白相对分子质量为66.0 kDa左右,与预测的结果相吻合。[结论]在MBP与目的蛋白之间引入了PPE酶切位点,可使MBP和SUVH6蛋白分离,并在使用MBP亲和层析和PPE酶切后,获得了纯度较高的SUVH6蛋白,表明成功构建了SUVH6-SRA-MBP融和蛋白原核表达系统。 相似文献
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无机有机肥磷配施对作物产量及土壤磷形态变化 总被引：2，自引：0，他引：2
采用盆栽模拟生物试验,研究等量施磷条件下无机有机肥磷不同配比处理对作物产量及土壤磷形态的影响.结果表明:无机肥磷:有机肥磷为80∶20的处理较100∶0处理前两季小麦和水稻产量略低,后两季小麦和大豆产量显著增高;80∶20处理的土壤活性有机磷和中等活性有机磷含量分别比100∶0处理增加60.0％和51.7％; 80:20处理的铁磷和闭蓄态磷含量较100∶0处理显著降低,铝磷和钙磷含量变化不大;80∶20和70∶30处理的土壤微生物生物量碳、磷含量均显著高于100∶0处理.因此,80∶20处理是本试验条件下无机有机肥磷配施的最佳比例,可为农业生产上提供借鉴. 相似文献
3.
Immunization of Male Mice with a New Recombinant GnRH Fusion Protein Reduces the Testicular Function 总被引：1，自引：0，他引：1
FANG Fu-gui YANG Ya-ping LIU Ya ZHANG Yun-hai TAO Yong WANG Suo-lu PU Yong ZHANG Xiao-rong 《中国农业科学(英文版)》2009,8(3):380-385
The objective of the present study was to evaluate the effect of an immunogenic maltose-binding protein-gonadotropin releasing hormone （GnRH-6-MBP） using genetic engineering. The synthetic mammalian tandem repeated GnRH hexamer gene was inserted into the expression plasmid pMAL-c2x. Recombinant GnRH-6-MBP protein was over- expressed in E.coli strain BL21. Amylose resin with affinity chromatograph was used to purify target protein. The reactiongenicity of fusion protein was identified by indirect enzyme linked immunosorbent assay （ELISA）, and the antigenicity and biological effects of GnRH-6-MBP were tested in mice. In the experiment, 20 male Kunming white mice of 20 d old were randomly divided into treatment and control group. Ten mice were immunized with 100 μg GnRH-6-MBP administered subcutaneously （s.c.） thrice at 2-week intervals with GnRH-6-MBP. Mice were sacrificed after 3 weeks following the booster injection, the testis was removed, weighed and measured, and the histological structure was observed. The reactiongenicity of fusion protein to GnRH antibody was much higher than the control. Active immunization against GnRH-6-MBP reduced remarkably （P 〈 0.01） the length and weight of the testis, and shortened the girth and width of the testis （P 〈 0.05）, and suppressed testicular spermatogenesis compared to the control mice. These results indicate that the recombinant GnRH-6-MBP acted as a strong immunogen and caused atrophy of the testis. 相似文献
4.
The competitive vs. complementary bacteria‐fungi interactions promote microbial release of Fe(III)‐fixed phosphorus: the roles of exogenous C application
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下载免费PDF全文 The phosphorus deficiency is very common in Fe(III)‐rich soil, and one of the key point is to clarify the condition in release or desorption of phosphorus from the Fe(III)‐rich minerals. The present study was to explore the effect of labile carbon on microbial reduction of Fe(III) and release of phosphorus in root‐free sub‐tropical soil. A two‐compartment microcosm was collected, in which the roots of Medicago sativa L. cultivar ‘Aohan' were confined within one compartment by a barrier of 30‐μm nylon mesh, while mycorrhizal hyphae could penetrate to the second compartment. Arbuscular mycorrhizal fungi (Funelliformis mosseae) were added to the root compartment and iron‐reducing bacteria (Klebsiella pneumoniae) were added to the hyphal compartment. Hyphal compartments were provided with two levels of additional carbon (0 and 23 mg C kg?1 soil as sodium acetate) and eight levels of inorganic phosphorus (0 to 35 mg P kg?1 soil as KH2PO4). At low phosphorus levels (< 5 mg P kg?1 soil), shoot biomass, and total biomass phosphorus were substantially less with added carbon in the presence of iron‐reducing bacteria. Carbon had little effect without iron‐reducing bacteria. At higher phosphorus levels (> 15 mg P kg?1 soil), the effect of added carbon was reversed; that is shoot biomass and total biomass phosphorus were greater with added carbon. Available soil phosphorus showed a similar response to added carbon—less at low levels of phosphorus and greater at higher levels of phosphorus. Microbial phosphorus in the presence of iron‐reducing bacteria was always higher with added carbon at all corresponding levels of soil phosphorus. Taken together, these results show that some phosphorus mobilized by iron‐reducing bacteria was converted into microbiological phosphorus, but there was an obligatory requirement for labile carbon for this to happen—reducing the amount of phosphorus that was absorbed by the mycorrhizal hyphae. Iron‐reducing bacteria and mycorrhizae showed a competitive interaction at lower levels of available soil phosphorus, and a complementary, or possibly a carbon‐dependent synergistic function at higher levels of available phosphorus. These results demonstrate that phosphorus released from ferralsols by iron‐reducing bacteria is positively mediated by both phosphorus and labile carbon and, hence, that phosphorus release and mobilization by iron‐reducing bacteria is likely to be enhanced by additions of exogenous carbon. 相似文献
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NF-YC9是模式植物拟南芥核因子Y家族成员之一,在开花调控等过程中发挥重要作用,但其蛋白的特性及其作用的分子机制尚待探究。本文首先扩增获得了NF-YC9基因的序列,采用重组技术构建了与麦芽糖结合蛋白MBP编码基因融合的重组表达载体,将其转化到大肠杆菌BL21中,进行了MBP-NF-YC9融合蛋白的诱导表达,并借助直链淀粉亲和层析介质纯化技术获得了电泳纯的MBP-NF-YC9重组蛋白,以备进一步对NF-YC9蛋白的生化特性进行全面检测。 相似文献
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三峡库区不同植被土壤微生物量碳氮磷生态化学计量特征 总被引：3，自引：0，他引：3
土壤微生物通过矿化和固持营养而显著影响土壤肥力。以湖北宜昌点军区三种植被类型(柏树、橘树、菜地)的土壤作为研究对象,对土壤微生物量碳氮磷及其生态化学计量进行了研究,探讨了不同植被覆盖土壤微生物量碳氮磷特征及它们之间的联系。结果表明:不同植被覆盖显著改变土壤微生物量碳氮磷含量,土壤微生物量碳(MBC)、氮(MBN)都是柏树地显著大于柑橘地和菜地,而菜地和柑橘地之间无显著性差异。土壤微生物量磷(MBP)的大小顺序为菜地柏树地橘树地。相关性分析的结果表明土壤有机碳、全氮和微生物量碳氮互相之间具有显著的正相关性(p0.01),土壤全磷与土壤微生物量磷之间也具有显著的正相关性(p0.01),这意味着土壤微生物量可以作为土壤肥力变化的指标。三种植被类型的土壤MBC/MBN之间都无显著性差异,而土壤MBC/MBP和MBN/MBP的变化顺序是橘树地柏树地菜地。MBC/MBP和MBN/MBP与MBP之间具有显著的负相关性(p0.01),而微生物碳氮与MBC/MBN,MBN/MBP和MBC/MBP之间都无显著的相关性。这意味着土壤MBC/MBP和MBN/MBP的变化主要与微生物量磷具有显著的关系。 相似文献
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四种蔬菜对DBP和DEHP及其代谢物的吸收累积研究 总被引：1，自引：0，他引：1
以不同浓度DBP和DEHP处理的土壤对青菜、菠菜、莴苣和萝卜进行盆栽实验,采用加速溶剂萃取,QuEChERS方法和硅烷衍生化对土壤和蔬菜样品进行前处理,结合气相色谱-串联质谱（GC-MS/MS）检测技术,分析了4种蔬菜中DBP、DEHP及其单酯代谢物MBP、MEHP从土壤中吸收的残留量分布规律和富集系数。结果表明,生长在高、低两个不同浓度DBP和DEHP土壤中的4种蔬菜都有DBP、DEHP及其代谢物MBP和MEHP的检出。萝卜中DBP和DEHP残留总量（Σ2PAEs）及其代谢物残留总量均明显高于其他3种蔬菜。4种蔬菜对土壤中的DEHP的生物富集因子BCF值都大于1,有明显的富集效应;而对于土壤中DBP有富集效应的则只有萝卜,其他3种蔬菜对DBP的BCF值都远小于1。相关研究结果表明,土壤中的DBP、DEHP能被蔬菜产品吸收富集,并且在体内代谢出毒性比母体更高的单酯产物。 相似文献
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A new insight into cystatin C contained in milk basic protein to bone metabolism: Effects on osteoclasts and osteoblastic MC3T3‐E1 cells in vitro 下载免费PDF全文
下载免费PDF全文 Takehiko Yasueda Yusuke Abe Mayumi Shiba Yoshie Kamo Yasuyuki Seto 《Animal Science Journal》2018,89(7):1027-1032
A milk protein fraction possessing alkaline isoelectric points (milk basic protein [MBP]) improves bone metabolism in vivo, and it inhibits bone resorption by osteoclasts and promotes mouse osteoblastic MC3T3‐E1 cells proliferation in vitro. Cystatin C (CysC) is a component of MBP and shows bone resorption inhibitory activity. Therefore, it is likely that MBP with higher CysC content improves bone metabolism more effectively. In this study, we prepared MBP with low and high contents of CysC and compared its effects on bone metabolism with standard MBP in vitro. Our results showed that the CysC content in MBP was positively related to not only bone resorption inhibitory activity but also MC3T3‐E1 cells proliferative activity. Furthermore, purified CysC stimulated MC3T3‐E1 cells proliferation. These results indicate that CysC contributes to promotion of MC3T3‐E1 cells proliferation, and MBP with higher CysC content shows enhanced bone resorption inhibitory activity and MC3T3‐E1 cells proliferative activity. CysC is considered an important factor in the effect on bone metabolism of MBP. 相似文献
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Tahereh Raiesi Ali Reza Hosseinpur 《Communications in Soil Science and Plant Analysis》2017,48(5):501-510
The objectives of this research were to evaluate the rhizospheric effects of bean (Phaseolus vulgaris) on phosphorus (P) availability and dissolved organic carbon (DOC), microbial biomass carbon (MBC), microbial biomass phosphorus (MBP), alkaline phosphatase (ALP) and P in the particulate fraction (PF-P) in some calcareous soils under rhizobox conditions. The results showed that DOC, MBC, MBP and ALP strongly increased in the rhizosphere soils compared with the bulk soils (p < 0.05). Also, the amounts of PF-P and P extracted with different tests in the rhizosphere were lower compared to the bulk soils (p < 0.05). The correlation studies showed that plant indices (dry yield and P uptake) had a positive relationship with Olsen-P, MBP, DOC, ALP and PF-P in both the rhizosphere and the bulk soil. Therefore, bean rhizosphere caused increases of DOC, MBC, MBP and ALP and decreases of available P and PF-P in the studied soils. In addition, the results of this research showed that the Olsen-P method and MBP and PF-P could be used to estimate bean-available P in the studied calcareous soils. 相似文献