Functions of poly-gamma-glutamic acid (γ-PGA) degradation genes in γ-PGA synthesis and cell morphology maintenance

Poly-γ-glutamic acid (γ-PGA) is an important biopolymer with greatly potential in industrial and medical applications. In the present study, we constructed a metabolically engineered glutamate-independent Bacillus amyloliquefaciens LL3 strain with considerable γ-PGA production, which was carried out by single, double, and triple markerless deletions of three degradation genes pgdS, ggt, and cwlO. The highest γ-PGA production (7.12 g/L) was obtained from the pgdS and cwlO double-deletion strain NK-pc, which was 93 % higher than that of wild-type LL3 strain (3.69 g/L). The triple-gene-deletion strain NK-pgc showed a 28 % decrease in γ-PGA production, leading to a yield of 2.69 g/L. Furthermore, the cell morphologies of the mutant strains were also characterized. The cell length of cwlO deletion strains NK-c and NK-pc was shorter than that of the wild-type strain, while the ggt deletion strains NK-g, NK-pg, NK-gc, and NK-pgc showed longer cell lengths. This is the first report concerning the markerless deletion of γ-PGA degradation genes to improve γ-PGA production in a glutamate-independent strain and the first observation that γ-glutamyltranspeptidase (encoded by ggt) could be involved in the inhibition of cell elongation.     

小麦TaMlo基因的原核表达、抗体制备及细胞化学分析(简报)

白粉病菌(Blumeria graminis)是一类高度专化性的寄生真菌,可侵染650多种单子叶植物和 9000多种双子叶植物,能够引起多种麦类作物的白粉病,给农业生产带来巨大的损失。由于白粉病菌生理小种多、变异快,所以利用专化性抗病基因难以解决植物的持久抗病性问题。人们在研究大麦白粉病时．发现大麦Mlo基因的隐性突变可导致大麦对绝大多数白粉病菌生理小种的高效持久的广谱抗病性。Schulze-Lefert等多家实验室合作于1997年成功克隆了野生的 Mlo基因。进一步研究表明．该基因编码一种植物特有的具有7个跨膜区和羧基端长尾的膜蛋白(Mlo),它可能对植物细胞的坏死起负调控作用。但Mlo基因如何表达及其在白粉病菌发育中的作用机制尚不清楚。     

1.42A crystal structure of mini-IGF-1(2): an analysis of the disulfide isomerization property and receptor binding property of IGF-1 based on the three-dimensional structure

Insulin and insulin-like growth factor 1 (IGF-1) share a homologous sequence, a similar three-dimensional structure and weakly overlapping biological activity, but IGF-1 folds into two thermodynamically stable disulfide isomers, while insulin folds into one unique stable tertiary structure. This is a very interesting phenomenon in which one amino acid sequence encodes two three-dimensional structures, and its molecular mechanism has remained unclear for a long time. In this study, the crystal structure of mini-IGF-1(2), a disulfide isomer of an artificial analog of IGF-1, was solved by the SAD/SIRAS method using our in-house X-ray source. Evidence was found in the structure showing that the intra-A-chain/domain disulfide bond of some molecules was broken; thus, it was proposed that disulfide isomerization begins with the breakdown of this disulfide bond. Furthermore, based on the structural comparison of IGF-1 and insulin, a new assumption was made that in insulin the several hydrogen bonds formed between the N-terminal region of the B-chain and the intra-A-chain disulfide region of the A-chain are the main reason for the stability of the intra-A-chain disulfide bond and for the prevention of disulfide isomerization, while Phe B1 and His B5 are very important for the formation of these hydrogen bonds. Moreover, the receptor binding property of IGF-1 was analyzed in detail based on the structural comparison of mini-IGF-1(2), native IGF-1, and small mini-IGF-1.     

荫蔽处理对不同耐荫性大豆GmPIF4s编码基因表达的影响

光敏色素互作因子(PIFs)是光信号响应的关键负调控因子,其中PIF4作为该家族核心成员在响应外界光温变化,整合多种激素合成和信号转导中起着关键性作用。为了明确大豆GmPIF4s ［phytochrome interaction factor 4s of Glycine max (L.) Merr.］ 的功能和编码基因对荫蔽诱导的表达响应,该研究以耐荫性大豆‘南豆25’(ND25)和不耐荫性大豆‘荣县冬豆’(RD)为材料,通过生物信息学和分子生物学等方法,比较分析了豆科植物PIF4s的生物信息学特点,并采用qRT PCR方法分析大豆不同组织(根、茎、叶、茎尖分生组织)、全光照和12 h/12 h光周期下不同荫蔽时间诱导叶片GmPIF4s及其不同耐荫性大豆材料间GmPIF4s的表达特征,为培育高产、耐荫的大豆品种提供理论依据。 结果表明：(1)在豆科植物中鉴定到53个同源PIF4s,其中大豆包含7个同源GmPIF4s,长度介于296~562氨基酸之间,分子量范围为41 491.9~62 228.39 Da,所有GmPIF4s均包含有bHLH结构域,无跨膜结构,为亲水性蛋白且定位于细胞核中;蛋白二级及三级结构预测显示,GmPIF4a、GmPIF4b与拟南芥AtPIF4结构最为相似。(2)qRT PCR分析表明,GmPIF4s基因表达具有组织特异性;在耐荫性大豆品种ND25中,GmPIF4a、GmPIF4b、GmPIF4d相对表达量在根、叶、茎和茎尖分生组织中表达最高,除GmPIF4a、GmPIF4e、GmPIF4f外,其余GmPIF4s在地上部各组织中表达量均相对较高;不耐荫性大豆品种RD中, GmPIF4a、GmPIF4b、GmPIF4c、GmPIF4d在叶中相对表量较高,除GmPIF4f和GmPIF4g外,其余GmPIF4s在地上部各组织中表达量均相对较高。(3)全光照下荫蔽处理时间延长,大豆GmPIF4a、GmPIF4b、GmPIF4c、GmPIF4d基因均受荫蔽的强烈诱导,表达量显著上调;12 h/12 h光周期条件下无论是正常光还是荫蔽条件下,随着处理时间的延长,白天GmPIF4s表达量呈现下调趋势,夜晚却呈现上调表达趋势,推测GmPIF4s受昼夜节律调控。(4)荫蔽条件下GmPIF4s在不同耐荫性大豆的表达显示,随着荫蔽时间的延长,耐荫性大豆品种ND25与不耐荫性大豆品种RD的GmPIF4a、GmPIF4b、GmPIF4c、GmPIF4d均上调表达,但在不耐荫性大豆品种RD中表达量相对更高,并于荫蔽处理8 h后两材料的GmPIF4s表达差异达到极显著水平。     

Recent advances in synthesis and surface modification of lanthanide-doped upconversion nanoparticles for biomedical applications

Lanthanide (Ln)-doped upconversion nanoparticles (UCNPs) with appropriate surface modification can be used for a wide range of biomedical applications such as bio-detection, cancer therapy, bio-labeling, fluorescence imaging, magnetic resonance imaging and drug delivery. The upconversion phenomenon exhibited by Ln-doped UCNPs renders them tremendous advantages in biological applications over other types of fluorescent materials (e.g., organic dyes, fluorescent proteins, gold nanoparticles, quantum dots, and luminescent transition metal complexes) for: (i) enhanced tissue penetration depths achieved by near-infrared (NIR) excitation; (ii) improved stability against photobleaching, photoblinking and photochemical degradation; (iii) non-photodamaging to DNA/RNA due to lower excitation light energy; (iv) lower cytotoxicity; and (v) higher detection sensitivity. Ln-doped UCNPs are therefore attracting increasing attentions in recent years. In this review, we present recent advances in the synthesis of Ln-doped UCNPs and their surface modification, as well as their emerging applications in biomedicine. The future prospects of Ln-doped UCNPs for biomedical applications are also discussed.     

环境污染物对巨噬细胞的影响及生物监测意义

随着工农业的发展,环境污染问题日趋严重。作为常见的环境污染物,二氧化硫经呼吸道转化为亚硫酸盐能引起多种呼吸系统疾病,并可能有促癌作用;重金属汞能引起中枢神经系统和多种器官损害,同时具有一定的免疫毒性,其危害已引起人们广泛关注。生物监测(bi0H10nitoring)是使用活     

不结球白菜Ty1-copia类逆转座子序列的克隆及表达

参照Ty1-copia类逆转座子逆转录酶的保守区设计简并引物,分别从10个不结球白菜(Brassica campestris ssp.chinensis)品种的全基因组中均扩增出260 bp左右的目标条带. 将目的条带回收、克隆和测序后进行分析,DNAstar分析发现,这些序列存在高度的异质性,28个核苷酸序列变化范围为224～278 bp,同源性范围为16.7%～83.0%.28条序列通过核苷酸聚类分为8个家族.推导氨基酸序列有移框突变、终止子突变或二者兼有;与已登录的不同物种同一类型逆转录酶氨基酸系统进化树分析表明,不结球白菜Ty1-copia类逆转座子与芥菜型油菜、拟南芥、芜菁、甜菜可能有共同的起源.半定量和实时定量PCR检测表明,水杨酸(salicylic acid)和Peronospora parasitica均能激活不结球白菜Ty1-copia类逆转座子,逆转座子在不结球白菜叶片中的表达特征说明它可能参与寄主对病原菌的抗性.     

基于GIS的浙北近海海域生态系统健康评价

在浙江省北部地区经济高速发展,科学高效利用海洋资源的同时,也不可避免的对浙北近海海域的生态系统造成一定的威胁。为了降低涉海工程对海洋生态环境可能造成的不利影响,通过分析浙江省北部海域高浊度、复杂水体等特点,利用层次分析法从水环境、沉积环境和海洋生物多样性构建了浙北近海海域生态系统评价指标体系,采用熵权法确定了相关因子的权重。在此基础上,综合GIS空间分析方法、数学模型,对2009年至2012年浙江省北部海域进行了实证研究。结果表明:(1)浙江省北部近海海域生态系统整体上处于不健康状态。含量较高的无机氮和磷酸盐及较低的海洋初级生产力是影响其环境质量的主要因素。(2)浙北近海海域生态系统主要薄弱区域与人类活动格局基本一致,由此推断,人类活动是影响浙北海域生态系统健康状态的主要因素。总体上,从2009至2012年浙北近海海域生态系统健康呈现逐年恶化的趋势。     

Interaction between GAMs and Mean Flow Shear During SMBI Injection into HL-2A Tokamak

Plasma Physics Reports - The interactions among geodesic acoustic modes (GAMs), mean flow, and mean flow shear were investigated using Langmuir probe arrays under periodic supersonic molecular beam...     

生物信息大分子功能的研究——酵母甘露聚糖对鼠Sarcoma-180瘤生长的抑制效应

酵母甘露聚糖（Mannan,简称Man）是能参与生物信息流影响生物体、特别是在糖基化方面起着重要调控作用的生物信息大分子.它是否能在抑制生物机体中肿瘤生长方面具有重要作用？研究结果表明：酵母甘露聚糖既能使患S-180瘤鼠的体质增强的同时又有抑制其体内所患S-180瘤生长的功效.Man抑制患鼠机体内的S-180瘤生长的功效（抑瘤率）随用量的增加而提高,当Man用量达360 mg（40 mg/Kg/d·9d）时,其抑瘤率达98.4％的最高水平,此时鼠体重增加1.66倍.Man的抑瘤功效有最佳适用量并存在性别敏感性,通常是雄性鼠的抑瘤率高于雌性鼠.Man抑制鼠S-180肿瘤生长的作用优于市售5-氟尿嘧啶的作用.