Bioinformatics Analysis of Bone Morphogenetic Protein 4 (BMP4) of Sheep

The assay was aimed to explore the biological characteristics of bone morphogenetic protein 4 (BMP4) of sheep,NCBI,DNAMAN DNAStar,TMHMM Server v.2.0,PsortⅡ,SignalP various bioinformatical softwares were used to speculate the physical and chemical properties,hydrophobic property,phosphorylation site,conservative structure domain,protein secondary structure of BMP4 protein.Also,the three-dimensional structure was forecasted with the SWISS-MODEL Workspace software.The results indicated that the BMP4 of sheep had high homologies with the BMP4 of various species.The encoded protein was a hydrophilic protein which was unstable.There was no transmembrane regions and it was likely to be located in the nucleus.What was more,there was signal peptide and eighteen phosphorylation sites.Through the forecast of functional domains,the protein had two functional domains,including the transforming growth factor-beta (TGF-beta) superfamily and TGF-beta propeptide superfamily.The result was consistent with the function of BMP4 gene family,it also demonstrated that BMP4 was a growth factor and it had the function of signal transduction.The amino acid homology between the predicted 3D structure of protein and template 3bmp.1.A was 88.29%.The bioinformatics analysis of BMP4 gene could provide reference for the further study in practice.     

基于pGhost4系统保加利亚乳杆菌乙酸激酶敲除载体的构建

保加利亚乳杆菌凭借其微生物特性和效能优异等特点成为当下产乳酸最重要的微生物菌株之一。乙酸是保加利亚乳杆菌代谢乳酸最主要的副产物,它的大量生成降低葡萄糖的代谢利用率,并且消耗了能量。乙酸激酶是控制乙酸生成的关键酶,敲除乙酸激酶基因,理论上可以阻断戊糖磷酸途径中乙酸的生成,从而优化代谢途径提高葡萄糖代谢乳酸的利用率。研究以Lactobacillus delbrueckii subsp.bulgaricus ATCC 11842公布的乙酸激酶基因ack序列设计引物,以保加利亚乳杆菌基因组DNA为模板,PCR克隆出ack上下游片段,利用重叠PCR技术将上下游片段拼接在一起,并连入具有温敏性的p Ghost4载体。     

Can additional N fertiliser ameliorate the elevated CO2‐induced depression in grain and tissue N concentrations of wheat on a high soil N background?

Elevated CO₂ stimulates crop yields but leads to lower tissue and grain nitrogen concentrations [N], raising concerns about grain quality in cereals. To test whether N fertiliser application above optimum growth requirements can alleviate the decline in tissue [N], wheat was grown in a Free Air CO₂ Enrichment facility in a low‐rainfall cropping system on high soil N. Crops were grown with and without addition of 50–60 kg N/ha in 12 growing environments created by supplemental irrigation and two sowing dates over 3 years. Elevated CO₂ increased yield and biomass (on average by 25%) and decreased biomass [N] (3%–9%) and grain [N] (5%). Nitrogen uptake was greater (20%) in crops grown under elevated CO₂. Additional N supply had no effect on yield and biomass, confirming high soil N. Small increases in [N] with N addition were insufficient to offset declines in grain [N] under elevated CO₂. Instead, N application increased the [N] in straw and decreased N harvest index. The results suggest that conventional addition of N does not mitigate grain [N] depression under elevated CO₂, and lend support to hypotheses that link decreases in crop [N] with biochemical limitations rather than N supply.     

苦瓜对白粉病的抗性与相关生理生化指标的关系

为探讨苦瓜抗白粉病的生理生化机制,以对白粉病不同抗性的4个苦瓜品系为材料,研究苗期感染白粉病菌后,苦瓜叶片生理生化指标的变化。结果表明:接种后,叶绿素和可溶性蛋白质量分数均呈先上升后降低的趋势;可溶性糖质量分数呈下降-上升-下降-上升的趋势;抗病品系的可溶性糖、叶绿素质量分数和过氧化物酶(POD)、多酚氧化酶(PPO)活性均高于或显著高于感病品系和高感品系;抗病品系的抗坏血酸(AsA)质量分数的上升和下降的幅度均小于感病品系和高感品系;接种后10~20d,抗坏血酸过氧化物酶(APX)活性表现为抗病品系感病品系高感品系。叶绿素、可溶性糖质量分数和POD、PPO活性与病情指数呈显著或极显著负相关。综上说明,白粉病菌侵染苦瓜后,抗病品系可通过保持较高的叶绿素质量分数,增加可溶性糖、可溶性蛋白、AsA质量分数及增强POD、PPO、APX活性来提高抗病性。叶绿素、可溶性糖质量分数和POD、PPO活性均可作为苦瓜对白粉病抗性早期鉴定的指标。     

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。     

不同基质活性炭对阿特拉津吸附特性研究

为了解决阿特拉津给土壤带来的污染问题以及寻找吸附效果较好的活性炭基质,本研究利用高效液相色谱(HPLC)技术检测了阿特拉津溶液和悬浮液经煤、木、果和竹质4种生物炭处理后的残留量,并对其在阿特拉津溶液和土壤中的吸附动态进行检测。结果表明,煤质活性炭对初始浓度为100 mg/L和10 mg/L的阿特拉津溶液和悬浮液的吸附效果较好,其吸附率为5.651%~68.42%;当阿特拉津初始浓度为100 mg/L时,煤质活性炭在40 min时对阿特拉津溶液的吸附率高达23.49%;与对照组相比,当阿特拉津的初始浓度为100 mg/L时,土壤中阿特拉津经煤质活性炭处理42天后的残留浓度最低,活性炭吸附率达到91.39%。综上,煤质活性炭能有效降解溶液及土壤中阿特拉津含量,这将为阿特拉津污染土壤的改良研究奠定基础。     

酸奶营养品质提升的加工工艺研究

牛奶中含有丰富的活性营养物质,包括乳铁蛋白、免疫球蛋白、乳过氧化物酶、α-乳白蛋白、β-乳球蛋白等。传统工艺生产的酸奶经过高温杀菌,活性营养物质几乎完全损失。为了避免这一情况,本文推出一种新的酸奶生产工艺,生产出的酸奶可保留大部分活性营养物质。首先将牛奶的乳脂进行分离,乳脂部分进行125 ℃、5 s杀菌,脱脂牛奶部分进行75 ℃、15 s杀菌,将两部分料液混合后加入菌种发酵至酸度75~85°T,破乳后冷却后熟8 h以上。经测定,新工艺酸奶中α-乳白蛋白、β-乳球蛋白、乳铁蛋白、免疫球蛋白、乳过氧化物酶含量分别为966 mg/L、1 963 mg/L、23.4 mg/L、102 mg/L、1 025 U/L,远高于传统工艺酸奶的68 mg/L、200 mg/L、0.7 mg/L、14 mg/L、0 U/L。通过对比新工艺与传统工艺酸奶保质期内的活菌数、黏度、酸度和口感,并无显著差异。     

三种融合标签对大熊猫轮状病毒结构蛋白VP6-VP7可溶性表达的影响

大熊猫轮状病毒(giant panda rotavirus,GPRV)是引起幼龄大熊猫腹泻的主要病原,对圈养大熊猫产生了较大的危害。轮状病毒结构蛋白VP6是一种载体蛋白,可介导黏膜免疫反应。VP7是轮状病毒结构蛋白中主要的中和抗原。因此,VP6-VP7的融合表达作为候选抗原对该病的防治具有重要的意义。传统大肠埃希菌原核表达存在表达量低、可溶性差以及纯度低等弊端。本研究使用醛缩酶(EDA)、谷胱甘肽S-转移酶(GST)、麦芽糖结合蛋白(MBP)3种融合标签,以实现获得表达量高和纯度高的GPRV-VP6-VP7重组表达蛋白。将扩增的VP6、VP7基因片段利用同源重组酶构建到含3种融合标签的表达载体pET21b上,将重组质粒转化至大肠埃希菌Rosetta(DE3)感受态细胞中进行低温诱导表达。用Ni-柱亲和层析法纯化目的蛋白,SDS-PAGE和Image J分析蛋白表达量和可溶性,Western blot分析得到表达的重组表达蛋白正确且具有蛋白活性。实验结果证明,EDA标签能显著促进VP6-VP7蛋白的原核可溶性表达,提高VP6-VP7蛋白表达量。