棉铃虫信息素结合蛋白2 cDNA的克隆、表达与组织特异性表达

 【目的】克隆、分析和原核表达编码棉铃虫Helicoverpa armigera (Hübner)信息素结合蛋白HarmPBP2的cDNA,并纯化得到HarmPBP2蛋白。【方法】以棉铃虫触角为材料,通过RT-PCR 技术以获得编码中棉铃虫信息素结合蛋白HarmPBP2 基因成熟蛋白开放阅读框序列,使用半定量RT- PCR对表达谱进行研究,并在使用pGEX-4T-1表达载体在BL21（DE3）系统中进行原核表达,使用GSTrap FF预装柱进行纯化并切去标签。【结果】克隆了命名为HarmPBP2（GenBank登录号：EU647241）的棉铃虫信息素结合蛋白基因,该序列全长457 bp,编码149 个氨基酸残基,前端是一段25个氨基酸长的信号肽,推测蛋白具有昆虫信息素结合蛋白典型的6 个保守的半胱氨酸残基的特征。棉铃虫HarmPBP2在雌雄成虫的触角、足和翅中都有表达,另外在雌虫下颚须也有表达。雌雄组织表达量顺序相同,触角中表达量最高,翅中次之,足中最少。雌虫组织中表达量高于雄虫。构建的原核表达载体pGEX/ HarmPBP2,在大肠杆菌BL21（DE3）中成功地表达出一个分子量约为40 kD的融合蛋白,经亲和层析纯化与酶切得到去标签的HarmPBP2蛋白。【结论】克隆、分析和表达了棉铃虫信息素结合蛋白HarmPBP2的cDNA序列,并对其进行纯化,为进一步研究其分子结构和功能奠定了基础,同时对其表达谱进行了研究。     

转谷氨酰胺酶对肉块冷黏结性能的影响

在对原料肉中添加 0 ,5 ,10g·kg-1转谷氨酰胺酶 ,分别与 0 ,10 ,2 0 ,30 ,4 0g·kg-1大豆分离蛋白、乳清浓缩蛋白、酪蛋白、明胶蛋白和卵清蛋白组成 5个试验组。在 4℃条件下催化反应 0～ 2 4h ,经成型、解冻后 ,用破碎样品时所做的功作为指标 ,测定不同试验组原料肉的冷黏结性能。结果显示 ,4℃、 2 0g·kg-1NaCl、 4 0g·kg-1大豆分离蛋白、 5g·kg-1转谷氨酰胺酶、反应时间 2～ 5h ,可以使原料肉获得较强的黏结能力 (2 4 2 3 5g·s)。转谷氨酰胺酶低温赋形性能的研究为原料肉的综合利用以及新型肉制品的开发提供了理论依据     

Growth of rumen papillae in weaned calves is associated with lower expression of insulin‐like growth factor‐binding proteins 2, 3, and 6

This study aimed to characterize the relationship between the growth of rumen papillae in calves and the mRNA expression of insulin‐like growth factor‐binding proteins (IGFBPs) in the rumen papillae. The length of rumen papillae, the mRNA expression of IGFBPs in rumen papillae by quantitative real‐time PCR, and the presence of insulin‐like growth factors I and II (IGF‐I and II) by immunohistochemistry (IHC) were analyzed in nine Holstein calves divided into three groups: suckling (2 weeks, n = 3), milk‐continued (8 weeks, n = 3), and weaned (8 weeks, n = 3). The length of rumen papillae was greater (p < 0.01) in weaned calves than in suckling and milk‐continued calves, whereas the expressions of IGFBP2, IGFBP3, and IGFBP6 genes were lower (p < 0.05) in the rumen papillae of weaned calves than in milk‐continued calves. Thus, rumen papillae length and IGFBP2, 3, and 6 expressions were negatively correlated. The IHC analysis showed that IGF‐I and IGF‐II were present in the rumen epithelium of calves. These results suggested that the growth of rumen papillae after weaning is associated with the induction of IGFs by the low levels of IGFBP2, IGFBP3, and IGFBP6.     

云南省玉溪市玉米致死性坏死病毒原的分子鉴定

玉米致死性坏死病是由玉米褪绿斑驳病毒(Maize chlorotic mottle virus,MCMV)和一种或多种马铃薯Y病毒科病毒复合侵染引起的。2014年1月在对云南省玉溪市玉米病毒病害的调查中发现了一些表现严重花叶、矮化叶片甚至整个植株坏死症状的玉米。对采集样品进行RT-PCR检测,所有样品中都同时检测到了MCMV和甘蔗花叶病毒(Sugarcane mosaic virus,SCMV),在一个样品中同时检测到了MCMV、SCMV和南方水稻黑条矮缩病毒(Southern rice black-streaked dwarf virus)。     

金雀异黄素对雌性大鼠体内促性腺激素及胰岛素样生长因子表达的影响

本试验旨在研究金雀异黄素(genistein,GEN)对雌性大鼠体内促性腺激素及胰岛素样生长因子表达的影响。选取40只SD雌性大鼠[体重(200±20)g],随机分为5组,分别为阴性对照(NC)组、GEN低(L)、中(M)、高剂量(H)组及阳性对照(PC)组,每组8只,NC组灌胃花生油(其他组灌胃试剂以此为溶剂);L、M、H组分别灌胃15、30、60 mg/(kg BW·d)GEN,PC组灌胃己烯雌酚0.5 mg/(kg BW·d)。试验期30 d。采用酶联免疫吸附试验(ELISA)法检测血清中卵泡刺激素(FSH)、黄体生成素(LH)、胰岛素样生长因子-1(IGF-1)、胰岛素样生长因子结合蛋白-1(IGFBP-1)含量;实时定量PCR法检测卵巢IGF-1、IGFBP-1 mRNA表达水平。结果表明:与NC组比较,试验组血清中FSH、LH含量有升高趋势,但差异不显著(P0.05),作用效果与PC组一致;试验组血清IGF-1含量略有降低,但差异不显著(P0.05),PC组显著降低(P0.05);试验组血清IGFBP-1含量显著或极显著升高(P0.05或P0.01),PC组显著升高(P0.05);试验组卵巢组织中IGF-1、IGFBP-1 mRNA表达水平均升高,其中M、H组显著升高(P0.05),与PC组变化一致。由此可见,GEN能够提高雌性大鼠血清FSH、LH含量、降低血清IGF-1含量、提高血清IGFBP-1含量,同时提高卵巢中IG FBP-1、IG F-1 mRNA的表达水平,这些指标协同作用于卵巢,能够促进卵泡的成熟,调节卵巢功能。     

Superfine grinding improves functional properties and antioxidant capacities of bran dietary fibre from Qingke (hull-less barley) grown in Qinghai-Tibet Plateau,China

The effect of particle size of hull-less barley (HLB) bran DF on antioxidant and physicochemical properties was investigated. HLB bran and extracted DF was ground by regular and superfine grinding, their particle sizes were determined using laser diffraction method. The results showed that superfine grinding could significantly pulverize DF particles to micro-scale; the particle size distribution was close to a Gaussian distribution. The soluble DF in HLB bran was increased effectively with superfine grinding. Insoluble DF with submicron scale showed increased total phenolic content (TPC), DPPH radical scavenging activity and ferric reducing antioxidant power (FRAP). With particle size reduction, the water retention capacity (WRC), swelling capacity (SC), oil binding capacity (OBC), and nitrite ion absorption capacity (NIAC) were significantly (p < 0.05) increased and the water holding capacity (WHC) had no significant change. A kind of health beneficial DF with higher soluble DF content, WRC, SC, OBC, NIAC and antioxidant activity was obtained using superfine grinding.     

The Distribution of Soluble Copper- and Zinc-Binding Proteins in Liver and Kidney of Chronic Copper-Poisoned Goats

The distribution of copper and zinc among the soluble proteins in the liver and kidney from chronic copper-poisoned goats was examined after gel filtration of the proteins. The concentrations of copper in the liver and kidney cortex from five experimentally copper-poisoned goats were: 550–810 µg/g liver and 190–420 µg/g kidney cortex (wet weight). In general the copper-binding proteins from, both the liver and kidney samples were separated into two different fractions with approximate molecular weights (m.w.) of > 65,000 and 10,000, respectively. From the liver samples, varying amounts of copper were eluted in a fourth fraction with m.w. < 2,000. In the majority of kidney samples the dominating copper-binding protein fraction was the high molecular weight fraction. Absolute amounts of copper recovered in the metallothionein-like protein fraction were nearly the samt for all samples investigated. The distribution of zinc-binding proteins in both liver and kidney samples was nearly the same. The high molecular weight fraction dominated, and no zinc was bound to metallothionein-like proteins.     

Isoflucypram,the first representative of a new succinate dehydrogenase inhibitor fungicide subclass: Its chemical discovery and unusual binding mode

Succinate dehydrogenase inhibitors (SDHIs) have played a crucial role in disease control to protect cereals as well as fruit and vegetables for more than a decade. Isoflucypram, the first representative of a newly installed subclass of SDHIs inside the Fungicide Resistance Action Committee (FRAC) family of complex II inhibitors, offers unparalleled long‐lasting efficacy against major foliar diseases in cereals. Herein we report the chemical optimization from early discovery towards isoflucypram and the first hypothesis of its altered binding mode in the ubiquinone binding site of succinate dehydrogenase. © 2020 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

烟粉虱内共生菌传毒相关groEL基因dsRNA载体的构建及其在烟草中的表达

烟粉虱内共生菌groEL基因编码一种与植物病毒传毒相关的GroEL蛋白.以B型烟粉虱为研究对象,利用1对特异引物,对B型烟粉虱体内的groEL基因进行PCR扩增,选取502 bp groEL基因片段构建dsRNA,并将其连接植物表达载体pBIN19,将重组质粒电激转化农杆菌菌株LBA4404.采用农杆菌浸染转化本氏烟草,PCR筛选表明,目的片段已整合至转基因植株的基因组DNA;RT-PCR结果表明,502 bp的groEL基因片段在转基因植株中被转录.     

金鱼Tgf2转座酶的原核表达及其DNA结合活性

金鱼Tgf2转座子基因属于Hobo/Activator/Tam3(hAT)超家族,其编码的活性转座酶能在多种鱼类转座中介导基因插入及诱变,因此在鱼类重要性状主控基因的筛选、功能解释及育种等方面具有重要的应用前景.鉴于Tgf2转座酶基因(JN886591)存在众多稀有密码子,本研究依据大肠杆菌对密码子的偏好性,对金鱼Tgf2转座酶基因进行密码子优化,所得Tgf2转座酶基因连接原核表达载体pET-28a(+),将重组载体转化到大肠杆菌BL21 (DE3)细胞.该大肠杆菌细胞在培养温度37℃、OD600≈0.5时用IPTG诱导获得高效表达,即菌体中含有8.8％的重组蛋白,上清液中含有4.0％重组蛋白.采用亲和层析纯化从菌体上清液中分离得到分子量约70 ku的可溶性重组蛋白,经MALDI-(TOF)/TOF串联质谱鉴定其为重组Tgf2转座酶.进而采用分子排阻色谱法研究转座酶的DNA结合活性,结果表明:所得重组Tgf2转座酶能识别并结合含Tgf2转座子特异性亚末端重复序列的DNA探针,即启动转座.采用原核表达体系高效获得重组Tgf2转座酶,为其作为工具酶应用于鱼类生物学研究奠定必要的基础.