农杆菌介导含硫氨基酸γ-zein转化菊苣的初步研究

含硫氨基酸具有动物营养与免疫相关的重要生理功能,为提高菊苣中含硫氨基酸含量,采用根癌农杆菌介导法将玉米种子贮藏蛋白含硫氨基酸基因γ-zein和绿色荧光蛋白GFP融合基因转入到菊苣无菌苗叶片中,经过共培养、潮霉素抗性筛选、分化、再生和炼苗,得到抗性植株。对抗性植株进行PCR、PCR-Southern、斑点杂交和RTPCR分析,结果表明,外源目的基因已经整合到菊苣基因组中并且得到了表达,为提高菊苣含硫氨基酸含量,改善其品质奠定了基础。     

Effect of Nutrient Restriction and Compensation on Weight,Serum Indexes and Gene Expression of Chest Fat Adipokines in Mongolian Lamb

To study the effect of nutrient restriction and compensation on weight,serum indexes and gene expression of chest fat adipokine in Mongolian lamb,thirty male lambs were randomly divided into control group (20.26 kg± 3.63 kg) and feed restricted group (21.29 kg±3.00 kg).The experiment lasted for 120 days,including 60 days for feed restriction and 60 days for compensatory feed.At the end of each stage,five lambs from each group were slaughtered and chest fat was collected.Serum concentration of triglycerides,non-esterified fatty acids (NEFA),total cholesterol were measured.Quantitative Real-time PCR were used to determine Adiponectin,Visfatin,PPAR-γ and ACC genes expression.The results showed that:①The lamb body weight of restricted group kept maintain level and that of control group showed a increase tendency.During the compensatory period,the average daily gain (ADG) of restricted group was significantly higher than control group (P< 0.05).②During the restriction period,serum triglycerides concentration of lambs in restricted group was significantly decreased at day 30 and 60 (P< 0.05),and NEFA concentration was significantly increased at day 30 (P< 0.05).After 60 days of nutritional compensation,all serum parameters we tested were not significantly different between restrict group and control group (P >0.05).③During the restriction period,PPAR-γ and ACC gene expression in restricted group were significantly lower than that of control group (P< 0.05),while no significant differences were found during the compensation period (P>0.05).Adiponectin and Visfatin genes expression in chest fat was not significantly changed throughout the experiment (P >0.05).We could know that nutritional restriction reduced fatty acid synthesis and adipocyte differentiation ability,adipose tissue was mobilized energy metabolism,and total cholesterol was recoveried growth on the compensation.However,the ACC and PPAR-γ genes in restricted group were significantly decreased with the body weight increased rapidly during the compensatory stage.Adiponectin and Visfatin were not significantly different between restricted and compensatory stages.     

归参汤对免疫抑制小鼠血清中IL-2、TNF-α和脾细胞Th1/Th2细胞因子的影响

为了探讨归参汤(GST)对免疫抑制小鼠血清中IL-2、TNF-α和脾细胞Th1/Th2细胞因子的影响。本试验采用环磷酰胺制造免疫抑制模型,利用双抗夹心ELISA法,观察归参汤对小鼠血清中IL-2、TNF-α的影响;RT-PCR法观察归参汤对小鼠脾细胞中INF-γ、IL-4、T-bet和GATA-3的mRNA表达的影响。结果显示:归参汤高剂量组可以显著增加免疫抑制小鼠血清中IL-2、TNF-α的含量(P<0.01),提高INF-γ的表达(P<0.05),降低IL-4的含量(P<0.05),升高T-bet(P<0.05),降低GATA-3的表达(P<0.05)。归参汤的免疫增强作用与升高血清中IL-2、TNF-α有关,且升高INF-γ/IL-4纠正免疫抑制小鼠Thl/Th2细胞因子的比例紊乱,其机制可能部分归因于对T-bet和GATA-3的mRNA表达的影响。     

60Co-γ和EMS诱变“天隆一号”突变体库变异特征的初步分析

为培育新的适应长江流域气候类型的大豆品种,利用60Co-γ射线和EMS分别对大豆天隆一号的种子进行诱变处理,构建大豆突变体库。对350份有表型变异的株系进行连续两年田间鉴定,运用60对SSR标记进行分子检测,并对籽粒表型变化明显的突变株进行苯基丙酸类代谢主要酶qPCR分析。结果表明:350份材料中有145份在株高、叶形、花色、种皮色、结荚习性等方面有稳定可见的表型变异,101份材料中检测到与野生型天隆一号存在至少1个SSR位点差异,其中M3-SD-1与M3-SD-2有超过10个标记的差异。SSR分析表明,表型发生变异的突变体主要是由DNA变化引起的,且突变体发生了多位点变异,突变位点也彼此不同。qPCR检测结果显示,种皮色、脐色等发生突变的3个材料苯基丙酸类代谢途径中关键基因的表达量发生显著变化。研究结果既可以为大豆品种改良提供新的种质资源,也有助于大豆功能基因组的研究。     

山羊IFN-γ基因的克隆表达与多克隆抗体制备

为获得山羊IFN-γ重组蛋白,提取山羊外周血淋巴细胞总 RNA,反转录得到 cDNA,以此为模板经PCR扩增获得IFN-γ的ORF区,构建重组克隆载体。经PCR扩增得到编码山羊IFN-γ成熟肽的基因序列,连入重组表达载体 pET-32a,转入 BL21（Codon Plus）感受态细胞中,测序并分析。重组表达载体经IPTG诱导后,对产物进行SDS-PAGE分析,并过镍柱纯化。用获得的纯化蛋白免疫家兔制备抗体。结果显示,编码山羊IFN-γ成熟肽部分的片段长432 bp;SDS-PAGE分析显示,融合蛋白大小约为34．9 ku,在30℃条件下,以0．3 mmol／L的IPTG诱导6 h,可得到大量可溶性表达的目的蛋白;间接 ELISA测定制得的多克隆抗体效价为1∶106。     

扶桑绵粉蚧辐照处理研究

为了寻求扶桑绵粉蚧有效除害处理方法,本文测定了γ-射线不同辐照剂量对该虫发育繁殖的影响,结果表明:随着辐照剂量的增加,各虫龄继续发育的比例明显降低;相同剂量水平下,各虫龄对辐照的耐受性随虫龄增大而增强,以3龄若虫和成虫的耐受性最强;大规模辐照测定表明,采用130Gy可有效阻止3龄若虫和雌成虫后代的发育繁殖。     

纳米Cs_2O/γ-Al_2O_3固体碱催化红麻籽油制备生物柴油

以纳米γ-Al2O3粉体为载体,应用等体积浸渍CH3COOCs制备Cs2O/γ-Al2O3催化剂,并通过TPD-CO2、XRD、TEM等手段对催化剂的碱性、结构和表面形貌进行表征,并将其用于催化红麻籽油制生物柴油反应.通过催化剂活性评价结果,分析了纳米固体超强碱制备过程及酯交换反应过程中各种因素的影响.结果表明,催化剂的粒径为10-25 nm,负载量为2mmol.g-1时,催化剂具有强碱性,其活性最好.甲醇与红麻籽油的摩尔比为9∶1,催化剂用量为油料的2.5%,反应时间3 h,转化率可达到90.7%.     

Increased Inflammatory Mediators in Horses Naturally Infected with Trypanosoma vivax. A Preliminary Study

The aim of this study was to measure the levels of inflammatory mediators in serum from horses naturally infected with Trypanosoma vivax. Banked serum samples collected during a previously reported T. vivax natural infection were used to analyze proinflammatory cytokines such as interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin 1 (IL-1), interleukin 6 (IL-6), and nitrite/nitrate (NO_x) levels. We evaluated 12 serum samples from horses from a farm in southern Brazil, four of which had parasitological and molecular diagnoses for T. vivax and presented with clinical signs of disease. Cytokines were assessed by quantitative sandwich enzyme-linked immunosorbent assay, and NO_x was measured using the modified Griess method. Levels of IFN-γ, TNF-α, IL-1, IL-6, and NO_x were increased in serum of infected animals compared to that in noninfected animals. Therefore, infection with T. vivax caused an increase in proinflammatory cytokines and nitric oxide content.     

微生物发酵合成γ-氨基丁酸的研究进展

γ-氨基丁酸是一种用于医药、食品中的新型功能性因子,由于化学合成法和植物富集法具有明显的缺点,因此微生物发酵合成γ-氨基丁酸成为目前研究的热点。该研究论述了产GABA的微生物、其培养基优化和诱变育种以及谷氨酸脱羧酶基因的研究进展。     

γ-射线辐照对植物保护性酶活性和MDA含量的影响

综述了在生长发育和储藏保鲜过程中不同剂量γ-射线处理对植物体内保护性酶活性和MDA含量的影响,并且在辐照保鲜中保护性酶活性和MDA含量的变化。