拟南芥At4g05060基因的酵母双杂交诱饵表达载体的构建、表达及自激活检测

为研究At4g05060基因功能,采用PCR扩增拟南芥VAMP(vesicle-associated membrane protein)家族基因At4g05060的ORF序列,定向克隆至酵母双杂交诱饵表达载体pGBKT7,经酶切鉴定并测序后将其转化至酵母Y187细胞;然后采用Western blotting技术分析At4g05060基因在酵母中的表达水平,通过缺陷型培养基培养进行自激活检测。结果表明,成功构建酵母诱饵表达载体pGBKT7-At4g05060,并在酵母细胞正确表达,表达产物对报告基因无自激活作用。     

Enrichment of cookies with glutathione by inactive yeast cells (Saccharomyces cerevisiae): Physicochemical and functional properties

Cookie is one of the favorite cereal products which could be formulated by using various ingredients for enrichment. The aim of this study was to determine the effects of the inactivate yeast cells addition to cookie formulation in terms of glutathione (GSH) content, antioxidant activities, physicochemical and sensorial properties. Also, the cookies were produced with GSH (pure, 98%) addition to compare the effects of inactive yeast GSH with pure GSH on revealed properties. According to results, The GSH contents and the antioxidant activities increased in dough and cookie samples with inactive yeast addition compared to control and pure GSH added dough and cookie samples. The increases in GSH contents and antioxidant activities were observed after baking in all samples. The pure GSH addition increased the moisture content, spread ratio and L* value while decreased the hardness and b* values of the cookies. The contrary results were obtained by inactive yeast addition in terms of the physicochemical properties where the protein content of the cookies increased with inactive yeast addition by approximately 25%. The loss of GSH content decreased while the antioxidant activities and nutritional value increased in cookies by inactivate yeast addition.     

利用酵母单杂交筛选拟南芥AtSTK基因表达调控蛋白的研究

拟南芥丝氨酸/苏氨酸蛋白激酶基因AtSTK的过量表达可以明显提高拟南芥的耐盐性。为进一步研究AtSTK基因表达的调控机制,以拟南芥基因组DNA为模板设计引物,扩增获得AtSTK基因的启动子序列,并构建到报告载体pAbAi,将重组报告载体质粒pAbAi-HYT利用BstbⅠ进行单酶切线性化,转化酵母菌株Y1H Gold,线性化的pAbAi-HYT整合到基因组中。然后,将纯化的拟南芥双链cDNA、pGADT7-Rec载体共转化含有报告载体pAbAi-HYT的酵母菌,将菌液涂布到含有100 ng/m L Ab A的SD/-Leu培养基上进行阳性克隆的酵母单杂交筛选。通过回复鉴定、序列测定,最终筛选获得了可能参与AtSTK基因表达调控的4个拟南芥基因。测序结果表明,酵母单杂交筛选获得的拟南芥基因AT3G32090含有WRKY转录因子保守结构域,为WRKY类转录因子的一员。因此,AtSTK基因的表达很可能接受MAPK信号传导途径中AT3G32090基因编码的WRKY类转录因子的调控,从而影响拟南芥植株的耐盐性。     

Fecal Gas Production of Ten Common Horse Feeds Supplemented With Saccharomyces cerevisiae

The study aimed to assess the nutritive value of 10 feeds (grains and forages) commonly used in horse nutrition in Mexico, on the basis of their chemical composition, in vitro organic matter digestibility (IVOMD) and in vitro gas production measurements with or without the supplementation of Saccharomyces cerevisiae (SC) at 4 mg/g DM. Fecal inoculum was obtained from 4 adult English Thoroughbred horses fed on restricted amount of concentrate and oat hay ad libitum. Substrates tested were: 6 concentrates (corn gluten meal, soybean meal, steam-rolled corn, steam-rolled barley, oat grain, and wheat bran) and 4 roughages (soybean hulls, corn stover, alfalfa hay, and oat hay). Gas production (GP) was recorded at 2, 4, 6, 8, 10, 12, 14, 24, 48, and 70 hours using the pressure transducer technique. Some ingredient × yeast interactions were observed (P ≤ .020) for the asymptotic GP and GP at 48 and 70 hours of incubation. Yeast addition increased (P < .001) the asymptotic GP of concentrates compared to roughages. Concentrate feeds had higher (P < .05) GP and lower (P < .001) rate of GP compared to roughages without yeast. From 24 to 70 hours of incubation, forages with or without yeast had lower (P < .05) GP compared to concentrates supplemented with SC. Forages had higher fermentation pH compared to concentrates but lower (P < .05) metabolizable energy, IVOMD, and microbial protein production compared to concentrates. Supplementation with SC increased (P < .05) the asymptotic GP of oat grain, soybean meal, soybean meal, steam-rolled barley, steam-rolled corn, wheat bran, corn stover, and oat hay, without affecting the rate of GP or lag time of oat grain, soybean meal, wheat bran, corn stover, and oat hay. Moreover, supplementation with SC increased (P < .05) metabolizable energy, IVOMD, and microbial protein production of steam-rolled barley, wheat bran, and corn stover, without affecting (P > .05) the fermentation of other feeds. Supplementation with SC improved fermentation of feeds with higher effects on concentrates compared to roughages. It was concluded that although SC mainly improves concentrate utilization by horses, it also improves fiber digestion when used on high-roughage diets fed to horses.     

酵母菌碱性磷酸酶的分离及部分性质研究

提取酵母菌中的碱性磷酸酶,并对其性质做出分析.结果显示酶促反应最适pH值为10.3,初速度 V0=0.01367μmol/L·min,以对硝基苯磷酸二钠(pNPP)为底物测得Km=0.535mmol/L、Vm=0.01429μmol/L.较低浓度的Mg2 对酶有较强的促进作用,2mmol/L时达到641%,磷酸氢二钠对碱性磷酸酶有竞争性抑制作用.     

益康XP酵母培养物在养殖业上的应用

就益康XP的组成、作用机理以及在畜禽和水产养殖中的应用情况进行了综述,表明益康XP在奶牛饲料中每天每头添加60g,日均产奶量增加;在仔猪和妊娠母猪饲料中分别添加0．25％和0．5％可提高仔猪的初生重和日增重;在蛋鸡饲料中添加0．1％或0．15％可提高其产蛋性能;在虾饲料中添加0．5％,可提高其产量和存活率。     

感染甘蔗花叶病毒甘蔗叶片cDNA文库的构建及评价

为了研究甘蔗(Saccharum officinarum L.)与甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)的互作机制,构建感染SCMV的甘蔗叶片的酵母双杂交cDNA文库。以热带种Badila为供试材料,提取感染SCMV植株的叶片总RNA,分离纯化获得mRNA,通过反转录反应合成3种读码框的双链cDNA。用Gateway技术,通过BP和LR反应,将cDNA迅速重组到酵母双杂交载体pGADT7-DEST上,构建酵母双杂交文库。结果表明,文库容量为2.0×107cfu,随机挑取24个克隆检测,重组率为100%,cDNA插入片段长度主要分布在1kb~2kb,文库质量较好。说明该文库的构建为筛选与SCMV互作的甘蔗寄主因子基因,研究甘蔗与SCMV的互作机制及甘蔗抗花叶病育种奠定基础。     

致病疫霉菌RXLR效应蛋白AVR1-like寄主靶标的酵母双杂交筛选

由卵菌病原物致病疫霉菌引起的马铃薯晚疫病是造成马铃薯毁灭性损失的病害。为了筛选致病疫霉菌RXLR效应蛋白AVR1-like(AL)的寄主靶标,以致病疫霉菌88069菌株的DNA为模板,克隆致病疫霉菌效应蛋白AL,并将其成功构建到pGBKT7载体上,形成诱饵载体。将pGADT7和重组诱饵载体共转化到酵母感受态AH109中,检测到诱饵载体无自激活作用。利用酵母双杂交技术以AL为诱饵筛选晚疫病菌侵染12 h后的番茄cDNA文库,初步获得6个候选AL寄主靶标。对6个候选靶标与AL进行一对一互作验证,结果表明其中有5个与诱饵载体强互作,1个弱互作。这些候选寄主靶标包括热休克蛋白同源蛋白、单脱氢抗坏血酸还原酶蛋白、过氧化物酶体蛋白等。     

Dietary supplementation of yeast cell wall improves the gastrointestinal development of weaned calves

The purpose of this study was to investigate the potential benefits of yeast cell wall (YCW) on the gastrointestinal development of weaned calves. Twenty healthy Holstein male calves (BW = 92 ± 8.29 kg and 60 ± 5 d of age) were randomly allocated into 2 groups: CON with no YCW, and YCW (accounted for 0.16% of the basal diet). The dietary concentrate-to-roughage ratio was 40:60. All the calves were fed regularly twice a day at 09:00 and 16:00 and had free access to water. The experiment lasted for 60 d. The results showed that calves fed YCW showed higher (P < 0.05) length, width, and surface area of papillae in the ventral sac of the rumen as compared to CON. For the dorsal sac of the rumen, the muscularis thickness was thicker (P < 0.05) in the YCW group when compared with CON group. The villus height of YCW calves was higher (P < 0.05) than that of CON in the ileum. Calves supplemented with YCW also showed a higher (P < 0.05) villus height-to-crypt depth ratio in the ileum. The YCW calves exhibited a greater (P < 0.05) thickness of the wall in the duodenum and jejunum. Calves supplemented with YCW improved (P < 0.05) the claudin 1 mRNA expression in the ileum and occludin mRNA expression in the jejunum and ileum. The YCW increased (P < 0.05) the contents of secretory immunoglobulin A in the jejunum and ileum of calves. In conclusion, dietary supplementation with YCW could improve the gastrointestinal development of weaned calves.     

利用载体微生物法生产血粉的研究

本试验用不同比例的各种原料,加入不同量的水制曲,来优选制曲配方,优选粗蛋白含量高的发酵血粉载体及鲜血加入量。研究用塑料袋法制做曲种,以饲养、代谢试验确定发酵血粉的饲用价值。 试验结果表明:a、最佳制曲配方为麸皮40％,胡麻饼60％,料水比为1:0.9;最佳发酵血粉载体配方为胡麻饼85％,麸皮15％,载体与牛鲜血比为1:1.15,发酵血粉产品含粗蛋白45.97％。b、塑料袋制曲不受外界湿度影响,杂菌不易侵入,曲种纯度高、成熟好,便于推广生产。C、用本试验的发酵血粉添加在日粮中喂鸡,饲养效果和对照组差异不显著,而经济效益比对照组高。