cDNA cloning and expression of erythropoietin in the plateau zokor （Myospalax baileyi） from the Qinghai-Tibet Plateau

All organisms respond to variation in their environments and manage environmental stress through metabolic adjustments.The plateau zokor(Myospalax baileyi) is an endemic and keystone subterranean rodent species that inhabits the Qinghai-Tibet Plateau between 2800 and 4200 m above sea level.It is a hypoxic-tolerant mammal with a high ratio of oxygen utilization that enables it to cope with its harsh surroundings.To explore the molecular mechanism of altitude acclimatization of the plateau zokor,we cloned the zokor erythropoietin(Epo) gene and used real-time PCR to compare Epo mRNA levels in zokors inhabiting 16 different altitudes.The full-length zokor Epo open reading frame was 579 bp that encoded a precursor peptide of 192 amino acids with a signal peptide of 26 residues.The Epo gene of the plateau zokor was 81%-95% homologous to that of human,mouse,rat,root vole and the Golan Heights blind mole rat,with the highest homology(95%) to species of the genus Spalax.Epo mRNA was detected mainly in the zokor kidney and spleen among 8 selected tissues.The level of Epo mRNA increased in the liver and kidney with increases in altitude.The increase in the kidney was 5 times that in the liver.Remarkably,expression of Epo mRNA in the kidney of zokors living at the highest altitude(4268 m) was 12-fold higher than that of zokors living at the lowest(2492 m) altitude.These findings provide essential information for understanding the possible role of Epo in adaptation to hypoxia in the plateau zokor.     

Cloning, sequencing and expression analysis of cDNA encoding a constitutive heat shock protein 70 （HSC70） in Fenneropenaeus chinensis

The cDNA encoding hsc70 of Chinese shrimp Fenneropenaeus chinensis was cloned from hepatopancreas by RT-PCR based on its EST sequence. The full length cDNA of 2090 bp contained an open reading frame of 1956 nucleotides and partial 5‘- and 3‘-untranslated region(5‘- and 3‘-UTR). PCR amplification and sequencing analysis showed the existence of introns in the region of 1--547 bp, but they did not exist in the region of 548--2090 bp of hsc70 cDNA. When the deduced 652 amino acid sequence of HSC70 was compared with the members of HSP70 family from other organisms, the results showed 85.9% similarity with HSC71 from Oncorhynchus mykiss and HSC70 from Homo sapiens. It also exhibited 85.8% similarity with HSP70 from Mus musculu and 85.4% with HSC70 from Manduca sexta. Expression analysis showed that hsc70 mRNA was espressed constitutively in hepatopancreas, muscle, eyestalks, haemocytes, heart, ovary, intestine and gills in Fenneropenaeus chinensis. No difference could be detected on hsc70 mRNA level in muscle between heat-shocked and control animals.     

Characterization of variation induced by low-energy N+ and cloning of differentially expressed cDNA of a mutant in Arabidopsis thaliana

Using Arabidopsis thaliana as experimental materials, the variations induced by low-energy N+ have been investigated. Germination rate of the treated seeds is lower than that of the control, and it decreases with the intensification of the radiation. The phenotypic variations have been observed in M2 plants irradiated with higher doses, such as chlorisis, semilethality, plant morphology, and changes of blooming habit and fertility. In random amplified polymorphic DNA (RAPD) analysis on M2 seedlings, some differences including band deletions or additions are found in treated plants compared to the control and the differences are associated with the radiation doses. One of the M1 plants from the seeds irradiated with the dose of 80×1015 N+/cm2 is a dwarf variant. Its stable M6 generation, mutant T80II, is used to construct subtractive cDNA library and to clone differentially expressed cDNA. A 721 bp cDNA fragment is partly homologous with GRF7 gene.     

Preparation,characterization, and antibacterial activity of γ-irradiated silver nanoparticles in aqueous gelatin

Colloidal silver nanoparticles (Ag-NPs) were obtained through γ-irradiation of aqueous solutions containing AgNO₃ and gelatin as a silver source and stabilizer, respectively. The absorbed dose of γ-irradiation influences the particle diameter of the Ag-NPs, as evidenced from surface plasmon resonance (SPR) and transmission electron microscopy (TEM) images. When the γ-irradiation dose was increased (from 2 to 50 kGy), the mean particle size was decreased continuously as a result of γ-induced Ag-NPs fragmentation. The antibacterial properties of the Ag-NPs were tested against Methicillinresistant Staphylococcus aureus (MRSA) (Gram-positive) and Pseudomonas aeruginosa (P.a) (Gram-negative) bacteria. This approach reveals that the γ-irradiation-mediated method is a promising simple route for synthesizing highly stable Ag-NPs in aqueous solutions with good antibacterial properties for different applications.     

Molecular cloning, expression and biochemical property analysis of AtKP1, a kinesin gene from Arabidopsis thaliana

Kinesins are common in a variety of eukaryotic cells with diverse functions. A cDNA encoding a member of the Kinesin-14B subfamily is obtained using 3＇-RACE technology and named AtKP1 （for Arabidopsis kinesin protein 1）. This cDNA has a maximum open reading frame of 3.3 kb encoding a polypeptide of 1087 aa. Protein domain analysis shows that AtKP1 contains the motor domain and the calponin homology domain in the central and amino-terminal regions, respectively. The carboxyl-terminal region with 202 aa residues is diverse from other known kinesins. Northern blot analysis shows that AtKP1 is widely expressed at a higher level in seedlings than in mature plants. 2808 bp of the AtKP1 promoter region is cloned and fused to GUS. GUS expression driven by the AtKP1 promoter region shows that AtKP1 is mainly expressed in vasculature of young organs and young leaf trichomes, indicating that AtKP1 may participate in the differentiation or development of Arabidopsis thaliana vascular bundles and trichomes. A truncated AtKP1 protein containing the putative motor domain is expressed in E. coil and affinity-purified. In vitro characterizations indicate that the polypeptide has nucleotide-dependent microtubule-binding ability and microtubule-stimulated ATPase activity.     

Characterization and expression of a cDNA, AmphiSDHD,encoding the amphioxus cytochrome b small subunit in mitochondrial succinate-ubiquinone oxidoreductase

In this study, an amphioxus cDNA, AmphiSDHD, encoding the cytochrome b small subunit in mitochondrial succinate-ubiquinone oxidoreductase, was isolated from the gut cDNA library of amphioxus Branchiostoma belcheri tsingtauense. It is 1429 bp in length, with an open reading frame of 465 bp coding for a protein of 154 amino acids. The deduced protein contains a mitochondrial targeting presequence of 65 amino acids rich in basic residues like arginine and hydroxy residues such as serine and threonine. Alignment of the amino acid sequences of AmphiSDHD and other eukaryotic SDHD proteins showed that AmphiSDHD has three transmembrane segments, and includes two histidine residues in the second transmembrane segment that are the putative binding sites for the heme b molecule. The phylogenetic tree constructed suggests that AmphiSDHD appears more closely related to vertebrate SDHD proteins than invertebrate ones. Northern blotting demonstrated that AmphiSDHD is ubiquitously expressed in amphioxus, being in line with the fact that SDHD is a house-keeping protein.     

Synthesis, characterization and antibacterial activity of copper, nickel and bimetallic Cu– Ni nanoparticles for potential use in dental materials

The antibacterial effect is a desirable property in dental materials.Development of simple methods for the preparation of nanosized metal particles has attracted significant attention because of their future applications due to unusual size-dependent antibacterial properties.Copper(Cu),Nickel(Ni) and bimetallic Cu–Ni nanoparticles were prepared by a simple chemical method and their antibacterial activity was tested against the widely used standard human pathogens Staphylococcus aureus(gram-negative) and Escherichia coli(gram-positive).Additionally,these nanoparticles were tested against the dental pathogen Streptococcus mutans.Our results are promising for potential use in dental materials science.     

Seasonal variation of gut Cyanophyta contents and liver GST expression of mud carp （Cirrhina molitorella） and Nile tilapia （Oreochromis niloticus） in the tropical Xiangang Reservoir （Huizhou, China）

Liver glutathione S-transferase(GST) plays a major role in the detoxification of microcystins(MCs) via conjugation to glutathione(GSH).We evaluated the relationship between seasonal variation in fish gut contents and the expression of GST isoforms in mud carp(Cirrhina molitorella) and Nile tilapia(Oreochromis niloticus).We quantified the abundance and diversity of plankton in the water column and foregut of mud carp and Nile tilapia in the tropical Xiangang Reservoir between October 2007 and July 2008.The mRNA expression of 7 liver GST isoforms was determined by real-time RT-PCR.The gut contents of both species were dependent on the amount and type of phytoplankton and zooplankton in the water.The expression of liver GST genes in Nile tilapia and mud carp was positively and negatively correlated,respectively,with the abundance of toxic cyanobacteria in the fore-gut.The expression of liver GST mRNA was correlated to the abundance of toxic cyanobacteria in the gut contents of both species,suggesting that mRNA expression of GST isoforms could be used as a biomarker in Nile tilapia and mud carp to monitor cyanobacteria blooms in reservoirs.     

Characterization and expression of a cDNA, AmphiSDHD , encoding the amphioxus cytochrome b small subunit in mitochondrial succinate ubiquinone oxidoreductase

Succinate ubiquinone oxidoreductase ,alsoknownascomplexII ,isthesmallestenzymecomplexfunctioninginboththetricarboxylicacidcycle(TAC)andtheaerobicrespiratorychainofeukaryoticcellmitochondriaandprokaryoticcells[1] .Itiscom posedoffournuclear encoded proteins ,SDHAorflavoprotein (70kD) ,SDHBoriron sulfurprotein(2 7kD) ,SDHCorCII 3(15kD)andSDHDorCII 4 (7— 9kD)inmosteukaryotes[2 ] .Thetwoproteins ,SDHAandSDHB ,comprisethesuccinatedehydroge nase (SDH) ,anenzymeoftheTACandaperipher…     

药材甲热激蛋白基因SpHsp60的克隆及温度胁迫下的表达

本研究采用RT-PCR技术克隆了药材甲热激蛋白60(Heat shock protein 60,Hsp60)基因的cDNA全长序列,命名为SpHsp60(GenBank登录号:KX778623).氨基酸序列分析表明SpHsp60具有Hsp60基因家族高度保守的基序,实时定量PCR技术检测了不同温度胁迫后该基因的表达量,结果表明:-5℃和0℃低温处理2h,药材甲成虫体内的SpHsp60的表达量均显著高于对照组,且42℃高温处理2h后的表达量也显著高于对照组.说明SpHSP60与药材甲应对极端温度胁迫相关.     

斑马鱼Cyclin C的cDNA克隆及其在发育过程中的表达

细胞周期蛋白C(cyclin C)与细胞周期依赖性蛋白激酶cdk8结合,通过磷酸化RNA 聚合酶 II 和 TFIIH调控转录.在脊椎动物胚胎发育过程中有关细胞周期蛋白C合子型转录激活机制尚知甚少.本研究克隆了斑马鱼细胞周期蛋白C基因,并用Northern杂交和整体原位杂交技术检测其在胚胎发育过程中的表达状态.结果显示,斑马鱼细胞周期蛋白C高度保守,与人细胞周期蛋白C有88％的蛋白序列同源;斑马鱼细胞周期蛋白C在母型期开始表达,其表达伴随中囊胚转换时期的合子型转录激活以及整个胚胎发育过程.     

Hydrothermal synthesis of pyrrhotite (Fex-1S) nanoplates and their antibacterial, cytotoxic activity study

The aim of this study was to synthesize and characterize Fe_(x-1)S 2D-nanostructures with pyrrhotite phase,as well as to explore their biological(antibacterial and cytotoxic)properties,namely the expression of reactive oxygen species(ROS)in the exposure of cells and bacteria.Based on hydrothermal synthesis,the characterization of asprepared 2D-nanostructures was performed by XRD,SEM,EDS,and TEM,in which the single-crystalline pyrrhotite phased Fe_(x-1)S nanoplate morphology was observed.The antibacterial activities of Fe_(x-1)S nanoplates against human pathogenic strains such as Staphylococcus aureus,Escherichia coli,and Enterococcus faecalis were tested.Minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)were determined following the broth microdilution method.Cytotoxicity and expression of intracellular ROS of pyrrhotite nanoplates on Human Gingival Fibroblast(HGF),Human Pulp Cells(HPC)and Human Osteoblast(HBC)were calculated.Cell viability was determined by the MTT method.All experiments were performed of three independent experiments and data were analyzed by Kruskal-Wallis and Mann-Whitney tests,also Pearson′s Correlation was performed.The nanoplates exhibited good bactericidal effect.All types of cells tested showed slight cytotoxicity.It was found that intracellular ROS is produced when cells and bacteria tested are exposed to pyrrhotite nanoplates in presence of both air and peroxide hydrogen.ROS production levels were higher in the bacteria than the cells exposed to these nanoplates.     

植物乳杆菌FZU122产细菌素的分离鉴定及其抑菌活性的研究

摘要：化学合成类食品防腐剂存在食品安全风险,乳酸菌细菌素是一类具有开发与应用前景的生物源食品抗菌剂。本研究以中国传统酸菜为原料,通过抑菌试验、生理生化鉴定与16 S rDNA测序技术筛选出一株产抑菌物质的植物乳杆菌FZU122。采用醇沉、葡聚糖凝胶色谱柱与HPLC分离、纯化并结合抑菌试验由该菌株发酵液中获得一种高纯度抗菌物质。基于纳升液相色谱-四极杆飞行时间串联质谱联用仪分析鉴定,该抗菌物质是一种由10个氨基酸组成的多肽ARLGLPVHVV,分子量1059.6553,带2个正电荷,将其命名为Plantaricin-fzu 122。抑菌实验结果显示Plantaricin-fzu 122对大肠杆菌、鼠伤沙门氏菌、耐甲氧西林金黄色葡萄等食源性致病菌具有不同程度的抑菌活性。综上所述,Plantaricin-fzu 122是一类可用于控制食源性致病菌污染且具有巨大开发潜力的生物防腐剂。     

亚硫酸盐在对虾保鲜加工中的残留变化

以南美白对虾为研究对象,研究常用的亚硫酸盐(亚硫酸钠和亚硫酸氢钠)不同浓度和不同浸泡时间处理对虾后,对虾在保藏过程中体内SO2含量的变化。结果表明:在同样浓度和处理条件下,亚硫酸氢钠比亚硫酸钠具有较好的防止对虾黑变的效果,对虾在冻藏过程中,体内SO2残留量逐渐消减。在对虾保鲜加工中,亚硫酸氢钠的最佳使用条件为:带头虾亚硫酸氢钠15 g/L,浸泡3 min;去头虾亚硫酸氢钠12.5 g/L,浸泡3min,该条件下能有效抑制对虾的黑变,虾体中SO2的残留量低于国标限量(0.1 m g/g)。     

Preparation,characterization and antibacterial property of naringin loaded PLGA nanospheres

Naringin is a predominant flavanone in grapefruit and shows a variety of biological effects such as antioxidative, anticancer, anti-inflammatory and antibacterial activity. However, its application in pharmaceutical field is limited by low water solubility, poor bioavailability and instability. To overcome the problem, naringin has been encapsulated in poly(lactic-co-glycolic acid) (PLGA) polymer by emulsion-diffusion-evaporation method in this work. Moreover, naringin loaded PLGA nanospheres were characterized by scanning electron microscope (SEM), dynamic light scatter method (DLS), fourier transform infrared (FTIR) spectra, UV–vis spectra and fluorescence spectra of DNA-EB competition displacement. The mean diameter of PLGA nanospheres and NRG/PLGA nanospheres was 123 ?± ?25 ?nm and 137 ?± ?30 ?nm, respectively. The drug encapsulation efficiency was 86.4% while the drug loading rate was 22.3%. The fluorescence spectra of the competitive DNA-binding experiments revealed that the functional activity of naringin was retained after loaded in PLGA. It is revealed that the initial burst effect happened in the initial 24 ?h and followed by sustained release lasting for 10 days. Moreover, the nanospheres exhibited strong antibacterial activity, and 99.9% of E. coli and S. aureus were killed when treated with naringin loaded PLGA nanospheres at the concentration of 0.2 ?mg ?mL^?1 within 24 ?h. Furthermore, the viable cells remained only 48% when the concentration of NRG/PLGA nanospheres was 32 ?μg ?mL^?1 and NRG/PLGA nanospheres was important for inhibition of cancer cells. It is concluded that the stable naringin loaded PLGA nanospheres could have potential application in food industry and nanomedicine field.     

苦荞蛋白磷酸酶2C家族的鉴定及表达分析

为了研究苦荞蛋白磷酸酶2C(PP2C)家族的成员和分类,及后续探究其在苦荞生长发育中的功能,本文利用生物信息学方法对苦荞PP2C家族进行鉴定、分类,并对其基因结构、保守基序、分子进化等进行分析.结果表明,苦荞PP2C家族有81个成员,划分为A-K的11个亚族,并且在同亚族中序列特征相似,而不同亚族间序列特征有一定差异;苦荞PP2C家族有14次基因重复事件.此外,qRT-PCR分析结果表明其A亚族基因在苦荞根、茎、叶、花、果中均有表达,除FtPP2C44外的8个基因在苦荞花、果中表达量较高;在苦荞幼苗中,除FtPP2C08外的8个基因均受ABA诱导表达量上调.上述结果揭示了苦荞PP2C家族的成员组成、序列特征、扩增和其A亚族基因的组织表达模式及受ABA诱导表达情况.     

凡纳滨对虾亲虾驯养、促熟、产卵及无节幼体培育

在水温27±2℃,盐度29±2;pH8.0~8.5,光强1 000 1x的条件下,对来自我国台湾的1 300对规格为35~45 g/尾的凡纳滨对虾亲虾进行为期73 d的驯养、促熟、产卵及无节幼体培育试验.结果表明,整个试验过程中,亲虾平均存活率为99.458%,波动区间[99.583%,99.333%],日产卵亲虾比例平均为3.506%,波动区间[3.89%,3.123%],雌虾单尾次日产无节幼体数量平均为18.928%,波动区间[19.691%,18.164%].无节幼体的日产量随繁殖驯养的延长,呈渐渐上升态势,从240万尾(第21天)至2 135万尾(第54天)平均为1 323万尾.此外,试验发现近成熟的雌虾也能接受交配行为.