An improved method for extraction of sorghum polymeric protein complexes     

《Journal of Cereal Science》2020

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.  相似文献   

抗凋亡融合蛋白PTD-Bcl-xL原核表达载体的构建及表达纯化     

王晓晔  石博妹  王英群  李珣  刘徳玉  李铭  李芳芳  胡传活 《华北农学报》2016,(1):1-7

人工抗凋亡蛋白PTD-Bcl-x L能保护多种因素引起的细胞异常凋亡,为了获得高纯度Bcl-x L与PTD(Protein transduction domains)的融合蛋白,首先采用TRIzol法提取SD大鼠肝脏总RNA,将RNA反转录为c DNA,设计引物以c DNA为模板,PCR扩增Bcl-x L基因,构建p UM19-T-Bcl-x L质粒,并对质粒双酶切鉴定和测序鉴定;其次设计包含PTD序列的Bcl-x L引物,以测序正确的p UM19-T-Bcl-x L质粒为模板,PCR扩增PTD-Bcl-x L序列,将扩增序列克隆入p ET28a载体,构建PTD-Bcl-x L蛋白的原核表达质粒p ET28a-PTD-Bcl-x L,并对p ET28a-PTD-Bcl-x L载体双酶切鉴定和测序鉴定;将p ET28a-PTD-Bcl-x L重组质粒转化大肠杆菌BL21(DE3),用IPTG诱导表达,并对IPTG诱导融合蛋白表达的浓度和诱导时间进行了优化;SDS-PAGE分析表达蛋白的可溶性情况,在变性条件下用Ni-NTA琼脂纯化融合蛋白;最后用SDS-PAGE、Western Blot及质谱对融合蛋白进行鉴定。结果表明:双酶切p UM19-T-Bcl-x L质粒出现约774 bp大小条带,p UM19-T-Bcl-x L质粒测序结果与NCBI数据库比对序列一致,表明成功构建p UM19-T-Bcl-x L质粒;双酶切p ET28a-PTD-Bcl-x L质粒出现约744 bp大小条带,p ET28a-PTD-Bcl-x L质粒测序结果与预期序列一致,表明成功构建了p ET28a-PTD-Bcl-x L原核表达载体;在IPTG诱导下p ET28a-Bcl-x L重组质粒在大肠杆菌BL21(DE3)中表达出36 k Da大小蛋白,最优IPTG诱导浓度为0.1 mmol/L,最佳IPTG诱导时间为6 h;SDS-PAGE电泳显示融合蛋白主要出现在菌液超声后的沉淀里,以包涵体形式表达,经Ni-NTA琼脂纯化获得了高纯度的融合蛋白;Western Blot和质谱鉴定证明IPTG诱导表达蛋白和纯化的融合蛋白为PTD-Bcl-x L蛋白。纯化得到了PTD-Bc L-x L融合蛋白,推进了PTD-Bcl-x L蛋白在猪、牛等家畜精液冷冻保存的应用进程。  相似文献   

用酶法制备藏羊血食用蛋白粉的研究     

胡勇  靳义超  吴海玥  仇桂芳  韩银仓  冶成君 《青海畜牧兽医杂志》2011,41(5):14-15

采用四因素三水平正交试验,对藏羊血制备食用蛋白的工艺进行了分析。经分析食用蛋白粉水解最佳条件为：酶解用酶量4.5g,酶解温度为46℃,酶解时间为7.5h,酶解的pH值为10.5,活性炭作用于水解液时由室温升至80℃所需最佳时间为6～8min。  相似文献   

Protein and energy concentrations of meat meal and meat and bone meal fed to pigs based on in vitro assays     

Hyunjun Choi  Chang Sik Won  Beob Gyun Kim 《动物营养（英文）》2021,7(1):252

The objective of this study was to develop equations for estimating ileal digestible crude protein (CP) and metabolizable energy (ME) contents of meat meal (MM) and meat and bone meal (MBM) as feed ingredients for pigs based on in vitro assays. Test ingredients were 4 sources of MM and 3 sources of MBM. Ash and CP contents of the ingredients ranged from 3.8% to 33.1% and 46.8% to 82.9% (as-is basis), respectively. In vitro ileal disappearance (IVID) of CP was determined and ileal digestible CP content was calculated by multiplying CP content by IVID of CP. In vitro total tract disappearance (IVTTD) of dry matter (DM) was determined and ME was calculated using gross energy, CP contents, and IVTTD of DM. The IVID of CP and IVTTD of DM ranged from 77.2% to 88.7% and from 82.7% to 92.4%, respectively. Calculated ileal digestible CP and ME contents ranged from 37.8% to 73.5% DM and 2,405 to 3,905 kcal/kg DM, respectively. Ash contents were negatively correlated (P < 0.001) with CP (r = −0.99), in vitro ileal digestible CP (r = −0.97), gross energy (r = −1.00), in vitro digestible energy (r = −0.97), and adjusted ME (r = −0.97). The most fitting equations for ileal digestible CP and adjusted ME were: ileal digestible CP (% DM) = 11.91 − 0.90 × Ash (% DM) + 0.74 × IVID of CP (%) (R² = 0.99) and adjusted ME (kcal/kg DM) = 130.85 − 50.90 × ash (% DM) + 47.06 × IVTTD of DM (%) (R² = 0.99). To validate the accuracy of the prediction equations for ME, mean bias and linear bias were determined using a regression analysis. Calculated ME values of MM and MBM were in a good agreement with data obtained from animal experiments based on a statistically insignificant bias in the models. In conclusion, ME concentrations of MM and MBM as swine feed ingredients can be calculated using ash concentration and in vitro disappearance of dry matter.  相似文献   

Comparative pharmacokinetics of marbofloxacin in healthy and Mannheimia haemolytica infected calves     

Ismail M  El-Kattan YA 《Research in veterinary science》2007,82(3):398-404

The pharmacokinetics of marbofloxacin were investigated in healthy (n=8) and Mannheimia haemolytica naturally infected (n=8) Simmental ruminant calves following intravenous (i.v.) and intramuscular (i.m.) administration of 2 mg kg(-1) body weight. The concentration of marbofloxacin in plasma was measured using high performance liquid chromatography with ultraviolet detection. Following i.v. administration of the drug, the elimination half-life (t(1/2 beta)) and mean residence time (MRT) were significantly longer in diseased calves (8.2h; 11.13 h) than in healthy ones (4.6 h; 6.1 h), respectively. The value of total body clearance (CL(B)) was larger in healthy calves (3 ml min(-1) kg(-1)) than in diseased ones (1.3 ml min(-1) kg(-1)). After single intramuscular (i.m.) administration of the drug, the elimination half-life, mean residence time (MRT) and maximum plasma concentration (C(max)) were higher in diseased calves (8.0, 12 h, 2.32 microg ml(-1)) than in healthy ones (4.7, 7.4 h, 1.4 microg ml(-1)), respectively. The plasma concentrations and AUC following administration of the drug by both routes were significantly higher in diseased calves than in healthy ones. Protein binding of Marbofloxacin was not significantly different in healthy and diseased calves. The mean value for MIC of marbofloxacin for M. haemolytica was 0.1+/-0.06 microg ml(-1). The C(max)/MIC and AUC(24)/MIC ratios were significantly higher in diseased calves (13.0-64.4 and 125-618 h) than in healthy calves (8-38.33 and 66.34-328 h). The obtained results for surrogate markers of antimicrobial activity (C(max)/MIC, AUC/MIC and T > or = MIC) indicate the excellent pharmacodynamic characteristics of the drug in diseased calves with M. haemolytica, which can be expected to optimize the clinical efficacy and minimize the development of resistance.  相似文献   

Nutritional Management of Cats and Dogs with Diabetes Mellitus     

《Veterinary Clinics of North America: Small Animal Practice》2023,53(3):657-674

  相似文献   

小麦杂种后代籽粒蛋白质含量的配合力研究   总被引：19，自引：1，他引：19  

金正勋  赵西华 《作物学报》1996,22(4):490-494

选用６个蛋白质含是高低不同的亲本，采用Ｇｒｉｆｆｉｎｇ双列杂交方法二，对亲本在Ｆ１－Ｆ３的蛋白质含量配合力作了研究。结果表明，蛋白质含量的ＧＣＡ与ＳＣＡ方差均极显著，表明在要试验中基因加性效应和非加性效应均起重要作用。但由于三个世代ｇｃａＭｓ／ｓｃａＭｓ值均极显著，且随着世代的推进这一比值又逐渐增加，因此蛋白质含量在杂种后代的表现主要还是由基因加性效应决定，随着世代的推进，基因加性效应越显重要，在  相似文献   

水肥耦合对冬小麦籽粒蛋白质及氨基酸含量的影响   总被引：5，自引：0，他引：5  

郑志松  王晨阳  牛俊义  张美微  张洁  姚宇卿 《中国生态农业学报》2011,19(4):788-793

为了解不同水、肥条件下小麦籽粒氨基酸及蛋白质含量的变化, 2009~2010 年度在河南省洛阳市农业科学院以“洛旱2 号”小麦为材料, 采用防雨棚池栽种植方式, 研究了不同灌水量、施氮和施磷量及其互作对小麦籽粒蛋白质及氨基酸含量的影响。结果表明: 不同灌水量和施氮量对小麦籽粒蛋白质和氨基酸均有极显著影响(P≤0.01), 且灌水×施氮互作效应显著(P≤0.05 或P≤0.01); 而施磷对其影响不显著。小麦籽粒蛋白质和氨基酸含量均随施氮量增加而增加, 随灌水量增加而降低, 但当灌水量超过282.0 mm、施氮量超过179.2 kg·hm^-2 时, 各指标的变化不再明显,蛋白质含量在高施氮量下略有下降。而必需氨基酸占总氨基酸含量的比例呈随灌水量增加而增加,随施氮量增加而降低的趋势。从不同水肥处理组合看, 蛋白质和氨基酸含量以处理组合N₁₀₅P₄₂W₁₂₇[施氮量105 kg(N)·hm^-2、施磷量42 kg(P₂O₅)·hm^-2 和全生育期灌水量127 mm, 下同]最高, 必需氨基酸/总氨基酸以处理组合N_30.8P₁₂₆W₂₈₂ 最高。籽粒产量随灌水量的增加而增加, 且产量高的处理其水分利用效率也较高。综合从蛋白质及氨基酸产量看, 以处理组合N_179.2P₁₂₆W₂₈₂ 表现最好, 即施氮量为179.2 kg·hm^-2、施磷量为126 kg·hm^-2、灌水量为282 mm。  相似文献   

玉米籽粒早期发育相关蛋白的差异表达特性     

于涛  李耕  张成芬  刘鹏  董树亭  张吉旺  赵斌 《作物学报》2017,43(4):608-619

玉米籽粒发育早期,代谢活动旺盛,细胞分裂与增大活跃,为后续贮藏物质的合成形成充足库容。为阐明籽粒早期发育的蛋白合成、积累与调控过程,本研究以夏玉米品种登海661为试验材料,在开花期人工饱和授粉后第3、第5、第10天取果穗中部籽粒,利用同位素标记相对定量(iTRAQ)技术分析其蛋白差异表达特性。玉米籽粒早期发育阶段总计鉴定及定量2639种蛋白,这些蛋白涉及多种生物过程与分子功能,其中代谢过程和分子过程是最主要的2个生物过程;催化活性和绑定功能是最主要的2个分子功能,这些生物过程与分子功能对籽粒早期发育具有重要作用。定量分析结果表明137种蛋白在籽粒发育早期显著差异表达,其功能涉及蛋白代谢、胁迫响应、细胞生长与分裂、碳水化合物与能量代谢、转运、次生物质代谢、淀粉合成、转录、油脂代谢、信号转导、氨基酸代谢等。其中,表达差异较大的是与蛋白代谢、胁迫响应、细胞生长与分裂以及碳水化合物与能量代谢相关的蛋白。表达模式聚类结果显示这些不同功能类别的蛋白协同表达,共同调控玉米籽粒的早期发育。  相似文献   

采用花粉管通道法将蛋白激酶基因导入玉米自交系的研究   总被引：4，自引：0，他引：4  

石薇  黄丛林  张秀海  吴忠义  杨德光 《华北农学报》2011,26(4):46-49

通过花粉管通道法将ZmPtil,ZmPtil-1,ZmCIPK2三种蛋白激酶基因植物表达载体分别导入玉米自交系吉444,经草丁膦筛选后得40株抗性植株,通过PCR检测其中28株为PCR阳性植株,平均转化率为1.33％,最后收获11株转基因植株.干旱条件下通过对转基因T1植株的单株籽粒产量、千粒重两个指标进行差异显著性和...  相似文献