H_2O_2在外源GA_4解除果梅季节性休眠中的信号作用

以GA_4处理的果梅休眠芽和转PmRGL2基因杨树叶片为材料,通过测定H_2O_2含量、抗氧化酶类活性及其编码基因和信号转导相关基因表达的变化,分析了H_2O_2在外源GA_4解除果梅休眠中的信号作用。结果表明:GA_4处理的果梅花芽的萌芽率显著高于对照,且H_2O_2含量在休眠解除时达到峰值,信号转导相关基因表达发生规律性变化;‘桃形梅’和‘丰后’的需冷量分别为574 CH(低温小时数Chilling hours)和1 108 CH,休眠解除前后,两品种叶芽中的H_2O_2含量没有显著差异,但需冷量低的‘桃形梅’具有更高的抗氧化酶类活性;转Pm RGL2杨树中NADPHoxi下调表达,有利于使H_2O_2含量维持在较低水平,并对赤霉素合成及信号转导相关基因的表达产生影响,而抗氧化酶类活性及其编码基因的表达量上升。推测GA_4通过调控抗氧化酶类活性而影响H_2O_2含量的变化,并引起上下游多种信号相关基因表达水平的变化,最终对休眠解除起作用。     

Screening of lentiviral vectors carrying effective siRNA targeting S1P2 gene

AIM:To screen the lentiviral vector carrying siRNA with higher efficiency of suppressing the sphingosine-1-phosphate receptor 2(S1P2) gene expression in the primarily cultured corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).METHODS:SHR and SD rats (n=5 each) were used for primarily culturing corpus cavernosum smooth muscle cells.The cells were randomly divided into 6 groups:SHR siRNA-1,SHR siRNA-2,SHR siRNA-3,SHR GFP,SHR control (SHR non-transfection group),and SD control (SD rat control group).Each group had 5 samples with 1.0×10⁵ cells of each sample.At 72 h after transfection (MOI=60) with lentiviral vectors carrying S1P2 siRNA into the SHR corpus cavernosum smooth muscle cells,the expression of GFP was observed under fluorescence microscope.The protein expression of S1P2,ROCK1,ROCK2 and eNOS in the corpus cavernosum smooth muscle cells,and the mRNA expression of S1P2,ROCK1 and ROCK2 were determined by by Western blot and RT-PCR.RESULTS:The transfection efficiency of the corpus cavernosum smooth muscle cells in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SHR GFP groups were>80%.Compared with SHR control group,the mRNA levels and the protein expression of S1P2,ROCK1 and ROCK2 in SHR GFP group showed no remarkable changes,while those in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SD control groups were significantly lower than those in SHR control group (P<0.05).The protein expression of eNOS in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SHR GFP groups were not significantly changed as compared with SHR control group,but that in SD control group was significantly higher than that in SHR control group.CONCLUSION:Three groups of siRNA lentiviral vectors targeting S1P2 inhibit the expression of S1P2 in the corpus cavernosum smooth muscle cells of SHR,and by silencing the S1P2 expression,the expression of ROCK1 and ROCK2 is inhibited.Among them,siRNA-1 has the highest inhibitory efficiency.     

剩余污泥臭氧化减量处置方法研究进展

综合臭氧污泥减量化近年来的发展和研究,对其技术原理和工艺应用进行分析,提出了其现存问题,并讨论了未来的技术发展万向.     

基于GWAS后分析策略研究TRAPPC9基因对奶牛产奶性状的遗传效应

本研究基于课题组前期对北京地区中国荷斯坦牛群体产奶性状全基因组关联分析结果,在新的中国荷斯坦牛群中将转运蛋白颗粒复合体9(TRAPPC9)基因作为产奶性状的候选基因进行遗传效应验证。研究利用混池测序寻找SNPs位点,结合前期GWAS结果筛选到的SNPs位点,进而分析这些多态性位点与中国荷斯坦牛产奶性状的相关性。结果发现,SNP1、SNP2位点对乳蛋白率和乳糖率效应均显著(P0.05);SNP3位点对乳脂率和乳蛋白率的效应均极显著(P0.01);SNP4位点对乳脂率有极显著影响(P0.01);SNP5位点对乳脂率和乳糖率的效应显著(P0.05)。同时发现TRAPPC9基因的表达量对产奶性状也有显著影响(P0.05),而TRAPPC9基因启动子区DNA甲基化对产奶性状无直接显著影响。该结果进一步表明,TRAPPC9基因可以考虑作为分子遗传标记应用于中国荷斯坦牛产奶性状的标记辅助选择。     

Suffer软件在构造面变化趋势研究中的应用

指出了枪马金矿位于河南灵宝小秦岭地区,其矿脉严格受断裂构造控制,构造产状沿走向发生改变和构造缓倾区是最有利的构造成矿位置.探讨了利用suffer软件的绘图功能,简单、快捷地识别和判断构造产状变化趋势和构造缓倾地段即有利于成矿部位.     

Immunogenicity of multiple-epitope antigen gene of HCV carried by novel biodegradable polymers

In order to develop a promising vaccine candidate utilizing a combined approach to induce both antibody production and T-cell activity, the DNA fragment containing MA of HCV with five conserved epitopes was synthesized. Two types of HCV vaccine candidates (the DNA type and DNA/polymers) were constructed using MA. PLA-PEG-PLA and PLGA-PEG-PLGA were synthesized and used as micelles with encapsulated plasmid pcDNA3.1(+)-MA. The preparation of copolymers, the cloning and analysis of recombinant plasmid DNA, in vitro expression, and immunogenicity in transgenic mice were evaluated in detail. The results indicated that even single immunization and oral immunization with DNA/polymers achieved satisfying immune responses in vivo tests. As biodegradable and nontoxic triblock copolymers, the novel copolymers demonstrated a great advantage, as they made long-term and single-immunizing vaccines possible; in addition, the copolymers showed a better adjuvant effect and scarcely any side effects.     

Development and application of a molecular marker for TMV resistance based on <Emphasis Type="Italic">N</Emphasis> gene in tobacco (<Emphasis Type="Italic">Nicotiana tabacum</Emphasis>)

Tobacco mosaic virus (TMV) caused serious loss in yield and quality of tobacco every year. It is a long-term goal to improve the tobacco resistance against TMV by tobacco breeding. N gene was the firstly reported TMV-resistant gene, which showed resistance against all Tobamoviruses except the Ob stain and belonged to the toll-interleukin-1 receptor/nucleotide-binding site/leucine-rich repeat class of plant resistance (R) genes. At present, N gene had already been widely used in tobacco conventional breeding, but there is rare available molecular maker used in marker-assisted selection of TMV resistance. In this study, we designed a pair of primers that specific amplify N gene fragment based on the sequence of N gene intron III, named N-marker. Then, we identified TMV resistance by two selecting methods, PCR with N-marker and inoculated with the TMV-C strain. Results from the two method showed that (1) 13 varieties among 67 tobacco varieties displayed hypersensitive reaction when inoculated with the TMV-C strain, also contained N gene fragments screened by PCR with N-marker; (2) 105 strains of 200 BC1 strains showed resistance against TMV when inoculated with TMV-C strain, meanwhile, 103 of the 105 strains contained N gene fragment verified by PCR with N-marker. Therefore, the N-marker is reliable for high throughput screening of germplasm resources and tobacco breeding materials in selection of N-mediated TMV resistance. Our study not only developed a molecular marker for tobacco breeding, but also identified new germplasm resources that are resistant to TMV.     

重金属污染土壤几种生物修复方式比较

土壤重金属污染是全球普遍存在的问题,生物修复因其环境友好且成本效益高而得到广泛关注。但不同生物修复技术有其优势和局限性,充分了解每种修复技术的特点,才能更经济、有效地对污染土壤进行修复。本研究阐述对比了目前的土壤重金属生物修复方法,包括植物修复（植物挥发、植物固定和植物提取）、转基因植物提取、螯合辅助植物修复、微生物辅助植物修复等技术的机制、优势、局限性和适用性等方面的差异。综述提出有效的生物修复技术需要土壤化学、植物生物学、遗传学、微生物学和环境工程等多学科的有机结合。根据污染土壤的特点,结合具有相应改良特性的转基因植物,是实现污染土壤大面积修复的有效方法。同时,农艺措施对天然超级积累植物的生物量和重金属提取能力的刺激作用还需要进一步挖掘。植物修复可以与其他几种传统修复技术有效结合,利用转基因技术建立土壤+植物+微生物的组合是未来修复技术发展的最佳途径。     

重组猪抑制素蛋白质中毒素残留检测及其理化稳定性

为探索纯化的重组猪抑制素蛋白质毒素残留情况以及重组猪抑制素蛋白质稳定性,本研究利用鲎试剂、气相色谱和SDS-PAGE方法对重组猪抑制素蛋白质中的细菌内毒素、β-巯基乙醇的残留量及其蛋白质在不同温度、极端p H值和反复冻融条件下的稳定性进行检测。结果显示,经过3次等电点沉淀洗涤后,重组猪抑制素蛋白质中细菌内毒素和β-巯基乙醇的残留量大幅减少。通过在不同温度下的处理发现,重组猪抑制素蛋白质在4℃环境下较稳定,可以长期保存;但在37℃时,稳定性快速下降,随着温度上升而降解加速。通过极端p H值和反复冻融试验发现,重组猪抑制素蛋白质对极端p H值和反复冻融具有较好的耐受性。     

接线错误造成变电一二次设备损坏事故

35 kV城东变投运于2012年,10kV为单母分段运行方式。10kV设备为中置式手车开关柜,2台主变各带一段母线,每段各有一组电压互感器(以下简称Ⅰ、Ⅱ母TV)。电压并列装置装设在Ⅰ母TV柜上,测量、计量电压采用开关柜柜顶小母线供电方式。