草胺膦抗性基因bar的原核表达与蛋白质纯化

通过PCR扩增,从pCAMBIA1300-bar质粒中获得bar基因编码区全长,经酶切鉴定和测序后证实,bar基因分离成功。用BamH和Hind酶切,将bar基因与原核表达载体pET28a(+)连接,构建成重组质粒pET28bar。将重组质粒pET28 bar转化大肠杆菌菌株BL21(DE3)后,获得高效表达。SDS-PAGE电泳分析显示,目的蛋白主要以包涵体形式存在,蛋白质分子量约为27 ku。将包涵体离心、洗涤和纯化后,得到高纯度的目的蛋白。该蛋白可以替代植物外源蛋白进行转bar基因产品的食用安全性检测,也可用于转bar基因产品ELISA检测的抗体制备。     

Protein to carbohydrate ratio in high-energy diets for Atlantic salmon (Salmo salar L.)

The effect of dietary protein to carbohydrate ratios (P/CH, weight percentage) of P34/CH21, P39/CH15 and P44/CH10 was studied in Atlantic salmon (Salmo salar L.) with respect to growth and feed conversion ratio (FCR) in one medium-scale experiment (Experiment 1) and to slaughter quality in two production scale experiments (Experiments 2A and 2B). The dietary fat was maintained at 290 g kg⁻¹ whereas the protein (fish meal) was exchanged with carbohydrate (wheat). Fish grown from approximately 1 to 4 kg were fed a restricted diet (iso-energetic on gross energy basis) or to satiation. Nitrogen, fat, starch and energy digestibilities were measured in a separate experiment. There were slight tendencies for lower growth (P = 0.06) and for higher FCRs (P = 0.06) in Experiment 1, and a slight tendency for a lower dress-out percentage in Experiments 2A and 2B (P = 0.10 and 0.20 respectively) with decreasing P/CH. The P/CH had no effect on the fat concentration of fillets, flesh colour or sexual maturation. The digestibility of starch decreased from 62.1% to 46.1% and the digestibility of energy from 84.9% to 79.5% when P/CH decreased from P44/CH10 to P34/CH21. Growth per unit digestible protein increased with decreasing P/CH. Feeding to satiation improved the growth but the FCR was higher than it was for restricted feeding. The calculated starch load per kg of fish growth increased with decreasing P/CH, whereas the effluent nitrogen decreased.     

与鸭C4结合蛋白互作的鸭疫里默菌外膜蛋白的筛选及鉴定

旨在筛选并鉴定与鸭C4结合蛋白（C4b-binding protein,C4BP）互作的鸭疫里默氏菌（Riemerella anatipestifer,R.anatipestifer）外膜蛋白。本研究将保存的R.anatipestifer复苏培养,提取外膜蛋白,以鸭C4BPα作为诱饵蛋白进行His pull-down及LC-MS/MS蛋白质谱鉴定,筛选与鸭C4BP可能发生互作的候选外膜蛋白;将各候选蛋白进行克隆、原核表达,免疫小鼠制备多克隆抗体,利用Far-western blot验证与鸭C4BP发生相互作用的R.anatipestifer外膜蛋白;针对候选蛋白及C4BPα各功能结构域进行克隆及原核表达,利用Far-western blot鉴定候选蛋白及与C4BP的相互作用位点;利用ELISA对补体因子C3b、C4b及C4BP在R.anatipestifer表面沉积情况进行测定,验证候选蛋白的功能。结果显示,经His pull-down及LC-MS/MS蛋白质谱分析,共筛选出3个与鸭C4BP发生相互作用的R.anatipestifer外膜蛋白,即ECE-1、SODs和Omp62;成功获得3个外膜蛋白多克隆抗体,ELISA检测3种多克隆抗体效价均超过1∶6 400,Western blot检测3种多克隆抗体可以与重组蛋白发生特异性反应;Far-western blot结果显示,仅ECE-1能够与C4BP发生相互作用,并且只有ECE-1全长能与鸭C4BP相互作用,而鸭C4BP与ECE-1的相互作用区域位于C4BPα的SCR 2和SCR 3;当健康鸭血清稀释度为3.125%时,ECE-1抗体能够显著促进补体因子C3b、C4b在R.anatipestifer表面的沉积作用（P<0.05）,当健康鸭血清稀释度为6.25%时,ECE-1抗体能够显著抑制C4BP在R.anatipestifer表面的沉积作用（P<0.05）。本研究成功筛选并鉴定出1个与C4BP互作的R.anatipestifer外膜蛋白ECE-1,为进一步阐明R.anatipestifer免疫逃逸机制奠定了基础。     

Effects of dietary protein to carbohydrate ratios on growth and body composition of juvenile yellow catfish, Pelteobagrus fulvidraco (Siluriformes, Bagridae, Pelteobagrus)

The present experiment was conducted to investigate the effects of dietary protein to carbohydrate ratios on growth and body composition of juvenile yellow catfish, Pelteobagrus fulvidraco . Nine diets were formulated to contain three protein levels (30%, 36% and 42%), each with three carbohydrate levels (24%, 30% and 36%). Each diet was randomly assigned to triplicate groups of 20 fish (initial mean body weight: 8.24±0.20 g) in indoor flow – through fibreglass tanks. The experiment continued for 8 weeks. Weight gain and specific growth rate were similar for the fish fed the 36% and 42% protein diets but higher than that fed the 30% protein diet. At the 36% protein level, carbohydrate contents varying from 24% to 36% ( P / E ratio of 24.0–28.2 mg protein kJ⁻¹) had no significant effects on growth performance and feed utilization ( P >0.05). Protein efficiency ratio tended to increase with dietary carbohydrate level at the same protein level. Dietary treatments significantly influenced body composition ( P <0.05), but not the condition factor, viscerosomatic index, hepatosomatic index and intraperitoneal fat ratio ( P >0.05). Based on these observations, 36% protein and 24–36% carbohydrate with the P / E ratio of 24.0–28.2 mg protein kJ⁻¹ seemed suitable for optimal growth and feed utilization, and carbohydrate could cause protein-sparing effect in diets for juvenile yellow catfish.     

Evaluation of pea proteins and poultry protein as fish meal alternatives in the diets for juvenile black sea bream,Acanthopagrus schlegelii

The potential of three different protein resources (pea protein isolate, PPI; pea protein concentrate, PPC; enzyme treated poultry protein, ETPP) as fish meal (FM) alternative protein in diets for juvenile black sea bream, Acanthopagrus schlegelii. (initial average weight 7.90 ± 0.13 g) was evaluated. Seven isonitrogenous and isoenergetic diets were formulated to replace FM at 0% (T0, control diet), 8% (designated as T1‐T3) and 16% (designated as T4‐T6) using PPI, PPC and ETPP respectively. Each diet was randomly assigned to triplicate groups of 25 juvenile fish for 8 weeks. At the end of the feeding period, survival rate was not significantly affected by dietary treatments. Growth performance in T6 (16% ETPP) group was significantly inferior to T0 group, however, weight gain and specific growth rate in other treatments showed no significant differences (P > 0.05). Mean feed intake, feed efficiency ratio and protein efficiency ratio were also poorer in fish fed in T6 than those of fish fed with the control diet respectively. Apparent digestibility coefficients (ADCs) of dry matter and crude protein for fish fed ETPP diets were significant lower than those of fish fed with the control diet, whereas ADCs of lipid were unaffected by dietary treatments. ADC's of dietary Leu, Ile, His and Lys was also significantly influenced. There were no marked variations in proximate compositions of dorsal muscle. With regard to plasma characteristics, significant difference was observed in triacylglycerol content. Ammonia concentration in plasma tended to increase in alternative protein diets as substitution level increased. There were significant differences in aspartate aminotransferase activities among groups, but alanine aminotransferase levels were unaffected by treatments. In conclusion, the present study demonstrated that PPI and PPC were potential protein sources for using in juvenile black sea bream diet. However, the substitution level of FM by ETPP should be limited within 16%.     

乳清分离蛋白—多糖乳状液制备与乳化稳定性研究

在pH值7.0时以乳清分离蛋白为乳化剂制备初级乳状液,利用静电自组装技术,通过加入具有不同电性的阴离子型果胶、中性瓜尔豆胶、阳离子型壳聚糖制备具有良好稳定性的乳清分离蛋白-多糖的二级乳状液,并考察pH值、乳清分离蛋白与多糖的比例对乳状液稳定性的影响.结果表明:瓜尔豆胶与乳清蛋白乳状液粒子间无静电作用;在pH值5.0 ～6.0条件下,壳聚糖能够吸附到乳状液粒子表面,但形成的乳状液稳定性较差;在pH值3.0～5.0、果胶质量分数为0.2％～0.5％条件下,乳清分离蛋白-果胶为乳化界面构建的二级乳状液,稳定性最高.     

三种酶制备牡蛎蛋白水解物的抗氧化活性和功能特性分析

目的：比较分析木瓜蛋白酶、胃蛋白酶和胰蛋白酶三种不同酶水解牡蛎获得的蛋白产物抗氧化活性和功能特性。方法：采用木瓜蛋白酶、胃蛋白酶和胰蛋白酶三种不同酶水解牡蛎蛋白获得水解物,并分别测定它们的在不同pH下的溶解度、乳化性以及不同浓度水解物的还原能力、DPPH自由基清除率和Fe^{2 +}螯合能力。结果：表明三种酶水解物都有较高的溶解度(p＜0.01),胰蛋白酶产生的水解物溶解高度达94.8％(p＜0.05);水解物溶解度和乳化活性指数在pH4显著降低(p＜0.05)。牡蛎蛋白质水解产物的还原能力、DPPH自由基清除能力和金属螯合活性随浓度的增加而增强,其中胰蛋白酶和胃蛋白酶抗氧化能力较强(p＜0.01)。结论：三种蛋白酶水解牡蛎蛋白可获得抗氧化活性强、溶解度高的水解物,并受浓度和pH的影响,它们的水解物可作为一种潜在的抗氧化功能多肽利用。     

Effect of cutting height and frequency on Leucaena leucocephala forage and wood production

Abstract

Leucaena leucocephala is a fast‐growing tree that can provide both high quality forage and firewood. The objective of this trial was to determine the optimum height and frequency of cutting for both wood and forage production. Cutting heights at 0.3 m, 0.6 m and 1.0 m were superimposed on 3‐month and 6‐month cutting frequencies on mature rows of L. leucocephala for three years. Effects of year or its interaction with the other factors were not significant (P≥0.10). There was an increase (P=0.07) in wood production but not forage production (P≥0.10) with the longer interval. Cutting height had an effect on forage (P<0.001) production, with 5.47 t ha^?1 a^?1 for the 0.3‐m height, 7.62 t ha^?1 a^?1 for the 0.7‐m height, and 8.71 t ha^?1 a^?1 for the 1.0‐m height. Cutting height also had an effect on wood production (P<0.001), with 7.22 t hd^?1 a^?1 for the 0.3‐m height, 9.33 t ha^?1 a^?1 for the 0.7‐mheight, and 11.55 t hd^?1 a^?1 for the 1.0‐m height. In this experiment, the six‐month interval produced more wood than the three‐month interval, but there were no differences in quantity of forage dry matter in L. leucocephala; there were advantages in both wood and forage production with the taller trunk base.     

50年黑龙江省大豆遗传改良的产量及品质变化

本研究针对黑龙江省过去50年推广的大豆品种产量及品质变化进行研究。结果表明,品种推广年代与产量之间表现显著的线性回归关系,产量呈逐年增加的趋势,平均每年提高1．27％。产量的提高与英数、英粒数、粒数及收获指数密切相关,而与百粒重关系不大。经过50年的遗传改良,大豆蛋白质和脂肪含量变化不大,且品种间变异幅度较小。R2期（盛花期）叶氮含量、单株叶氮量、比叶氮及LAI（叶面积指数）与产量及品质关系不大,但此时期干物质重与产量里负相关关系,生育前期的干物质积累对产量的贡献不大。     

Effects of Supplementing a Basal Diet of Chloris gayana Hay with One of Three Protein-rich Legume Hays of Cassia rotundifolia,Lablab purpureus and Macroptilium atropurpureum Forage on Some Nutritional Parameters in Goats

Growth and digestibility experiments were conducted on growing East African type goats offered Chloris gayana hay supplemented with one of three high-protein (119–128 g CP/kg DM) legume hays, Cassia rotundifolia (cassia), Lablab purpureus (lablab) or Macroptilium atropurpureum (siratro), and crushed maize to investigate the feed intake, digestibility, growth and urinary excretion of purine derivatives. Goats in the supplemented groups had higher total dry matter and nitrogen intakes and higher N retention and body mass gains than unsupplemented counterparts. The digestibility of dry matter, organic matter and neutral detergent fibre were increased by protein supplementation. Animals on supplemented diets had higher fractional outflow rates of particulate matter from the rumen. The production of protein by ruminal microbes and the efficiency of microbial N production were increased by supplementation. It was concluded that a mixture of low-quality grass hay (61.9 CP/kg DM) and either cassia, lablab or siratro hay, and maize grain can provide a productive balanced diet for growing goats.