日粮不同CP及RUP水平对奶牛瘤胃发酵的影响

试验旨在研究不同水平的粗蛋白（CP）及瘤胃非降解蛋白（RUP）的日粮对奶牛瘤胃代谢的影响。试验选择18头健康、体重相近的荷斯坦奶牛作为试验动物,随机分为6组,每组3头。采用2×3因子完全随机试验设计,其中CP设7.5%（低）、9.0%（中）、10.5%（高）3个水平,RUP设45%CP（低）、55%CP（高）2个水平,共6个处理组,饲喂相同基础日粮,精粗比为3∶7,预饲期10d,饲喂后2、4、8、12、24h从食道采集瘤胃液2d,测定瘤胃液pH值,NH3-N、微生物蛋白质（BCP）、挥发性脂肪酸（VFA）含量。结果表明,日粮CP水平对瘤胃液NH3-N浓度的影响极显著（P〈0.01）,对BCP及VFA的含量影响较小,总体而言,9.0%CP水平组对奶牛的瘤胃发酵略好于其他两组;日粮中高RUP（55%CP）可以显著降低瘤胃pH值且有降低NH3-N浓度的趋势,同时,对瘤胃BCP及VFA的合成有一定的促进作用。该试验条件下,日粮中CP（9.0%）和RUP（55%CP）可在一定程度上改善瘤胃代谢。     

猪链球菌蛋白表面展示系统的构建及展示蛋白的初步观察

为构建猪链球菌(Streptococcus suis)蛋白表面展示系统,本研究通过序列分析,确定猪链球菌的LPxTG蛋白及其信号肽(SP)和胞壁锚定基序(CWA),通过PCR扩增Peno-SP、GFP、CWA的DNA片段并融合,构建强启动子Peno控制表达编码SP-GFP-CWA融合蛋白的DNA片段,将该重组DNA片段连接pSET2载体,获得蛋白表面展示质粒,转化猪链球菌,构建得到以GFP为报告蛋白的猪链球菌蛋白表面展示系统。结果显示,利用猪链球菌的10个LPxTG蛋白及其SP和CWA序列,构建了10个含有Peno-SP-GFP-CWA融合片段的重组pSET2表面蛋白展示质粒pSsPSD1至pSsPSD10,分别转化猪链球菌05ZYH33,PCR鉴定显示其中7个转化猪链球菌。采用western blot初步检测其展示蛋白,结果显示,7个转化阳性菌株均能有效表达GFP蛋白,以成熟GFP条带为指标,均表现出了一定的外源GFP表面展示水平,分别命名为SsPSD1、SsPSD2、SsPSD4、SsPSD7-SsPSD10,其中SsPSD1、SsPSD4、SsPSD8和SsPSD9表面展示水平相对较好,在猪链球菌表面展示外源蛋白方面具有很好的潜力。本研究首次尝试建立猪链球菌蛋白表面展示系统,为猪链球菌表面递呈外源蛋白或抗原提供了新的策略。     

Effect of energy and protein levels on performance,egg quality,and nutrient digestibility of laying pigeons

A total of 3,240 female Taisheng pigeons at 40 wk age were fed 9 diets containing 3 ME levels (2,630, 2,770, and 2,940 kcal/kg) and 3 CP levels (14.0, 15.0, and 16.0%) in a factorial arrangement. These diets were fed to investigate the effects of ME and CP on performance, egg quality, and nutrient digestibility. Female pigeons were housed under the same managerial conditions and randomly assigned into 9 treatments (6 replicates of 60 birds each). Two female–female paired pigeons housed in one cage were under a 16L:8D lighting cycle. Feed in pellet form and water were provided for ad libitum consumption. The study lasted 12 wk. As dietary ME level increased from 2,630 to 2,940 kcal/kg, BW, egg production, and apparent DM, crude fat (CF), and phosphorus (i.e., P) digestibilities increased (P < 0.05), feed intake decreased (P < 0.05), and feed conversion efficiency increased (P < 0.05). Increasing the level of dietary protein from 14 to 16% resulted in an increased (P < 0.05) BW, shell percentage, length of the major axis, and DM digestibility. Therefore, dietary ME had a greater influence on the performance of Taisheng laying pigeons than did dietary protein.     

Cloning and Protein Structure Analysis of Hypoxia Inducible Factor-1α Gene in Yak

In this study,hypoxia inducible factor-1α(HIF-1α) gene was cloned from yak spleen by RT-PCR,and it was divided into two sections for amplification.The sequence and protein structure were analyzed using related bioinformatics tools.The length of the coding region was 2 472 bp,which encoded 823 amino acids.Its molecule mass was 92.13 ku,and PI was 5.09.It displayed as a hydrophilic protein,whose secondary structure was mainly composed of random coil and α-helix.It had 69 phosphorylation sites and 4 N-glycosylation sites.Phylogenetic tree displayed that Maiwa yak,Bos grunniens,Bos mutus and Bos taurus gathered into one cluster.Maiwa yak HIF-1α gene was successfully cloned in this study,it provided a viable reference for further study of genetic characteristics of HIF-1α.     

菠菜新品种蔬菠2号的选育

蔬菠2号是以雌性系221为母本、自交系213为父本配置的菠菜一代杂交种。该品种植株直立,生长势强,叶片大、深绿色、尖圆形,叶面平展;植株生长速度较快,适于早春露地、秋露地及设施栽培。     

An improved method for extraction of sorghum polymeric protein complexes

A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm.     

重组减蛋综合征病毒KnobS干酪乳杆菌的构建与表达

以干酪乳杆菌为传递载体,以开发食品级安全的减蛋综合征病毒疫苗为目标,构建了一个温度敏感型自杀型质粒系统p ORZP-UKD。将该质粒电转化到干酪乳杆菌L.casei中,经过2次升温处理和5-氟尿嘧啶抗性筛选,挑选阳性克隆,采用PCR和SDS-PAGE方法进行鉴定。KnobS基因整合到干酪乳酸杆菌基因组内,并实现融合蛋白KnobS的分泌表达。表明得到了一株具有无任何选择性标记、携带有KnobS表达盒元件的基因组整合的重组干酪乳酸杆菌KnobSΔupp L.casei,且整合的外源基因能够随着细菌的复制而进行表达,为减蛋综合征病毒免疫保护性抗原KnobS疫苗的进一步研制提供了试验数据。     

白藜芦醇通过激活去乙酰化酶1/腺苷一磷酸激活的蛋白激酶信号通路影响牛脂肪细胞凋亡

本试验旨在研究植物提取物白藜芦醇(RES)对牛皮下脂肪细胞凋亡率以及去乙酰化酶1(SIRT1)/腺苷一磷酸激活的蛋白激酶(AMPK)信号通路关键因子的mRNA和蛋白质表达量的影响。选取18月龄鲁西黄牛的皮下前体脂肪细胞,在细胞分化的第0天,更换为RES浓度分别为0(对照)、100、200和400μmol/L的培养液处理48 h,每组设3个重复。应用Hoechst33342染色检测细胞凋亡的形态学变化,流式细胞分析仪检测细胞凋亡率,荧光定量PCR(q PCR)和免疫印迹试验(Western-blot)分别检测SIRT1/AMPK信号通路上的关键因子SIRT1、A MPKα、叉头转录因子1(Fox O1)、B细胞淋巴瘤/白血病-2(Bcl-2)、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、促凋亡蛋白Bcl-2相关X蛋白(Bax)的mRNA和蛋白质表达量,并利用油红O染色鉴定脂肪细胞。结果表明,与对照组相比:不同浓度RES处理后的牛皮下脂肪细胞凋亡率均极显著增高(P0.01);不同浓度RES处理后的SIRT1、AMPKα、caspase-3、Bax的mRNA表达量均显著或极显著提高(P0.05或P0.01),Bcl-2的mRNA表达量极显著降低(P0.01);不同浓度RES处理后的SIRT1、AMPKα、caspase-3的蛋白质表达量均显著或极显著提高(P0.05或P0.01);200和400μmol/L RES处理后,Fox O1的mRNA和蛋白质表达量显著或极显著提高(P0.05或P0.01),Bax的蛋白质表达量极显著提高(P0.01),Bcl-2的蛋白质表达量极显著降低(P0.01)。100μmol/L RES处理后时,Fox O1的mRNA和蛋白质表达量以及Bcl-2、Bax的蛋白质表达量均与对照组差异不显著(P0.05)。由此可见,RES通过激活SIRT1/AM PK信号通路,同时激活通路下游的Fox O1,促进了牛皮下脂肪细胞的凋亡,为通过营养调控技术降低牛皮下脂肪沉积提供了一定的理论基础。