Site-specific uv crosslinking of minihelix DNA and TrpRS fromBacillus subtilis

In order to investigate the recognition mechanism and the relationship between structure and function of minihelix DNA with Tryptophanyl-tRNA Synthetase (TrpRS), TrpRS from Bacillus Subtilis was purified. Four minihelix DNAs were chemically synthesized and the photoreactive reagent s⁴T was incorporated into three of them at the positions of G73, T72 and T55 corresponding to tRNA^Trp. The apparatus for uv crosslinking was devised and the parameters for uv crosslinking were optimized. The results indicated that the G73 and T72 base of minihelix DNA interacted with TrpRS directly. The uv crosslinking reaction was improved by the dose of uv irradiation and the concentration of both TrpRS and minihelix DNA.     

Anticodon and acceptor stem nucleotides in tRNA(Gln) are major recognition elements for E. coli glutaminyl-tRNA synthetase

The correct attachment of amino acids to their corresponding (cognate) transfer RNA catalysed by aminoacyl-tRNA synthetases is a key factor in ensuring the fidelity of protein biosynthesis. Previous studies have demonstrated that the interaction of Escherichia coli tRNA(Gln) with glutaminyl-tRNA synthetase (GlnRS) provides an excellent system to study this highly specific recognition process, also referred to as 'tRNA identity'. Accurate acylation of tRNA depends mainly on two principles: a set of nucleotides in the tRNA molecule (identity elements) responsible for proper discrimination by aminoacyl-tRNA synthetases and competition between different synthetases for tRNAs. Elements of glutamine identity are located in the anticodon and in the acceptor stem region, including the discriminator base. We report here the production of more than 20 tRNA(2Gln) mutants at positions likely to be involved in tRNA discrimination by the enzyme. Unmodified tRNA, containing the wild-type anticodon and U or G at its 5'-terminus, can be aminocylated by GlnRS with similar kinetic parameters to native tRNA(2Gln). By in vitro aminoacylation the mutant tRNAs showed decreases of up to 3 x 10(5)-fold in the specificity constant (kcat/KM)14 with the major contribution of kcat. Despite these large changes, some of these mutant tRNAs are efficient amber suppressors in vivo. Our results show that strong elements for glutamine identity reside in the anticodon region and in positions 2 and 3 of the acceptor stem, and that the contribution of different identity elements to the overall discrimination varies significantly. We discuss our data in the light of the crystal structure of the GlnRS:tRNA(Gln) complex.     

Accurate localization and excision of genomic islands in four strains of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> and <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis>

Mobile genomic islands (GIs) can be excised from the chromosome, then form a circular intermediate and be reintegrated into the chromosome by the GI internal integrase. Some mobile GIs can also be transferred into a new receptor cell by transformation, conjugation, or transduction. The action sites of the integrase are usually flanked direct repeats (DRs) of the GIs. Accurate localization of the flanking sequences is a precondition for determining the mobility of the GI. Mobile GIs are generally associated with transfer RNAs (tRNAs). Based on the correlation between flanking sequences and tRNA sequences, the flanking sequences of 11 putative mobile GIs in Pseudomonas aeruginosa PAO1, P. aeruginosa PA14, P. fluorescens Pf-5 and P. fluorescens Pf0-1 were identified. Among the 11 GIs, Pf0-1GI-1 is responsible for benzoate degradation. PAO1GI-1, Pf5GI-2, Pf5GI-3, and Pf5GI-4 were confirmed experimentally to be excised from a chromosome to form a circular intermediate. The action sites of the integrases are these GIs direct repeats. Due to distinct DRs, cutting sites for the internal integrase of PAO1GI-1, Pf5GI-2, Pf5GI-3 and Pf5GI-4 were determined outside the T-loop of the tRNA^Gly gene, outside the anticodon loop of the tRNA^Ser gene and tRNA^Lys gene, and at the asymmetric 3′-end of the tRNA^Leu gene, respectively. PAO1GI-1 and other mobile GIs may be transferred into many different strains that belong to different phyla because of the clear flanking sequences. This study describes basic information about the action sites of the integrases, assesses the mobility of GIs, and can help design and transfer mobile GIs to candidate strains.     

Transfer of bacterial blight resistance from <Emphasis Type="Italic">Oryza meyeriana</Emphasis> to <Emphasis Type="Italic">O. sativa</Emphasis> L. by asymmetric somatic hybridization

Asymmetric somatic hybrid plants were produced between cultivated rice (Oryza sativa L.) and wild species [O. meyeriana (Zoll. etMor, exSteud.)] with high resistance to rice bacterial blight. X-ray-irradiated protoplasts of the wild species were used as donor and chemically fused with iodoacetamide-inactivated protoplasts of rice cv. 02428 to produce hybrids. Seventy-two plants were regenerated from 623 calli based on metabolic complementation. The morphological characters of the plants closely resembled that of the rice. Simple sequence repeats were employed to identify their hybridity. Cytological analysis of root-tips revealed that their chromosome number varied in the range of 27--38. The somatic hybrids were inoculated with strains of Xanthamonas oryzae pv. oryzae at adult growth stage and demonstrated the resistance to bacterial blight introgression from the O. meyeriana.     

拟南芥雄性不育突变体opw(only pollen wall)候选突变基因的克隆

在T-DNA插入突变体Salk_059463株系的群体中,筛选到两株雄性不育突变体,对TDNA序列上的一对引物进行PCR鉴定,结果表明:其基因组中没有T-DNA插入.遗传分析表明这两株雄性不育突变体由同一单个隐性基因控制,引起不育的主要原因是从花药发育的第8期开始,小孢子细胞质内容物逐渐减少直至消失,到花药发育的第12期,药室内的小孢子只剩下一个花粉壁空壳,故该突变体命名为opw(only pollen wall).利用图位克隆的方法对OPW基因进行了定位,结果表明OPW基因位于第二条染色体上分子标记T28M21和T3G21之间的12 kb区间内,该区间内一共有21个基因注释.通过克隆区间内的基因并测序发现opw-1突变体基因组中At2g40140基因编码序列的外显子在第289和第290个碱基之间插入了一个A碱基,而opw-2突变体基因组中At2g40140基因编码序列的外显子在第412和第413个碱基之间插入了一个T碱基,造成的编码序列移码使第424至第426碱基成为终止密码子,故At2g40140是编码OPW的候选基因.     

An improved method for measuring the stability of a three-state unfolding protein

In the current three-state protein unfolding model, the two transitions are considered to be independent and each transition is fitted to a two-state unfolding model. This three-state unfolding process is therefore composed of two sequential two-state unfolding processes. In this paper, a modified method is presented to determine the value of the unfolding free energy [δG _total⁰(H₂O)] for the three-state unfolding equilibrium of proteins. This method is demonstrated on the apoCopC protein mutant, Y79W-W83F-Cu, which unfolds via a three-state process. The value of ΔG _total⁰(H₂O) calculated using the modified method was found to be more accurate in determining ΔG _total⁰(H₂O) than the previously reported method.     

食药同源植物火麻的研究进展及开发策略

食药两用植物火麻(Cannabis sativa L. subsp. sativa)具有优良的植物油(火麻油)和植物蛋白(火麻蛋白),全身是宝,在食品、医药和工业用品行业具有巨大的市场开发潜力。随着大健康产业的发展,火麻蛋白的系列产品必将越来越深入我们的生活。国内外对火麻的研究报道比较丰富、研究方向多,但是缺少全面系统的总结分析。本文通过查阅国内外对火麻研究的文献报道,综述食药两用植物火麻的最新研究进展,找出火麻发展的不足并提出开发策略:要实现火麻产业经济,需要建立有优质的种质资源圃,健全高产优质栽培技术及火麻加工、销售体系,并对其化学成分和营养生理功能机制深入研究,才能促进火麻产业化、规模化和可持续化的发展。     

Taxonomic relationship betweenOryza minuta andO. officinalis based on AP-PCR analysis

The arbitrarily primed polymerase chain reaction (AP-PCR) technique was used to detect polymorphisms in genomic DNA fromOryza minuta, O. officinalis, and related rice species. The results show that the genetic distance betweenO. minuta andO. officinalis is much higher than that between two known species,O. sativa andO. rufipogon, showing thatO. minuta andO. officinalis should be classified as two different species in genusOryza, and that the AP-PCR technique can be used as a powerful tool for studying taxonomic relationships among related species, and even subspecies. Supported by the National Natural Science Foundation of China Yi Qingming: born in Apr. 1938, Professor     

Resistance of oat to ‘take-all’ causing fungus (Gaeumannomyces graminis var.tritici)

Evaluation with the pot assay at seedling stage in greenhouse showed that oat (Avena sativa) was highly resistant to take-all disease to which, however, wheat (Triticum aestivum) was extremely susceptible. The oat roots were shown to be inhibitory to the invasion and spread of take-all causing fungus G. graminis var. tritici by the following criteria: (ⅰ) less infection sites were observed (about 1/7 of those in wheat); (ⅱ) the ectotrophic growth of G. graminis var. tritici on oat roots was much slower than that on those of wheat, and the runner hyphae appeared as kidney- or fork-shaped hyphopodia on the surface of oat roots which could not be discerned on that of wheat roots; (ⅲ) the period from inoculation to penetration into the epidermis of oat roots was about 2.9 times as long as that of wheat; (ⅳ) the infection hyphae were hindered substantially when it was about to penetrate into the epidermis of oat roots with the mycelium deformed; and (ⅴ) the cortical layer of oat roots was revealed to be unsuitable for the G. graminis var. tritici infestation as some lysed hyphae were found therein, and the spread of hyphae from the first layer of cortex to the pericycle needed 108 h, about 1.8 times as long as it did on wheat roots.     

Effect of kinase-negativeEGFR gene on differentiation of embryonic germ cell line EG4

EG4 cells derived from primordial germ cells (PGCs) of 10.5 d post coitum 129/svJ mouse embryos can be used as a model system for in vitro differentiation study due to their pluripotential development ability. EG4 cell lines with stable expression of kinase-negative EGFR cDNA, designated EG4-EGFR^d, were generated by gene transfection. We found that: (ⅰ) EG4-EGFR^d cells share the similar morphology and growing character with wildtype cells that can maintain undifferentiated state in long term culture. (ⅱ) Treatment of EG4 cells with RA resulted in differentiation of adipocyte, while in mutant clones of EG4-EGFR^d, adipocytes were sparse or absent under the same condition, indicating the role of EGFR expressed during adipocyte development. (ⅲ) Histological analysis showed that predominant tissues in teratocarcinomas derived from EG4-EGFR^d cells and wildtype cells are different. A large amount of undifferentiated cells was present in those coming from mutant cell clones. In addition some cardiac and skeletal muscles are prominently differentiated cell types. EG4 wildtype cells produced multiple differentiated cell types of three primary germ layers such as cartilage, epithelia and neural tube. These studies suggested that EGFR-dependent differentiation was inhibited in kinase-negative EG4 cells.     

Cloning and Characterization of the fecC Gene Necessary for Optimal Growth under Iron-Deficiency Conditions in the Cyanobacterium Anabaena sp. PCC 7120

The fecC gene encoding a putative iron (Ⅲ) dicitrate transporte rwas cloned from nitrogen-fixing cyanobacterium Anabaena sp. PCC 7120, and inactivated. The mutant grows normally in medium with NO3^- , NH1^- or without combined nitrogen. But in iron-deficient medium, the mutant grows slowly. Photosynthetic properties were compared between the mutant and the wildtype strain, the content of photosynthetic pigments in the mutant is lower than that of the wild-type. The results of RT-PCR experiments show that the fecC gene is expressed under iron-deficient conditions, but is not expressed under iron-replete conditions. These results revealed that fecC gene product is required for optimal growth under iron-deficient conditions in Anabaena sp. PCC 7120.     

Prescribing symmetric scalar curvature onS 2

The problem of prescribing scalar curvature in S2 is discussed, and the solvability of the e-quation - Δu + 2-Reu = 0 on S2, is given, where R∈C0(S2). It is known that there are some obstructions . Some new results are given by seeking a solution of the minimax type. For example, supposing that R is G symmetric and is constant on the set of fixed points on S2 under G (where G is a subgroup of O(3)), It is proved that the equation is solvable if and only if R is positive somewhere.     

The high-resolution climate recorded in the δ18O ofPorites lutea from the Nansha Islands of China

A Porites lutea core from Yongshu Reef of Nansha Islands covering 50 years growth history was analyzed for oxygen isotopic composition with monthly and seasonally resolution. The calibration of the δ ¹⁸O with the instrumental temperature indicated that the coral δ ¹⁸O is a good indicator for sea surface temperature (SST) and air temperature ( t ). It can be used to reconstruct the SST and air temperature of the Yongshu Reef sea area. In addition, the coral δ ¹⁸O provides signatures for the intensity of the East Asia monsoon and it is a record for the activities of El Niño events. With the calibrated SST and air temperature formulas, the most recent fifty years SST and air temperature were reconstructed based on the coral δ ¹⁸O, thus back up the understanding of the climate of Nansha Islands to 1950, far beyond the limit of the instrumental recording since September 1988. It was found that, in general, increasing 1℃ air temperature results in 0.24‰ decrease in skeletal δ ¹⁸O.     

基于生成对抗网络的人脸图像彩色化方法

提出了一种新的基于生成对抗网络的人脸图像彩色化方法.所提出的网络结构包含两组生成对抗子网络,每个子网络由一个生成器和判别器组成.其中,一个对抗子网络A（包含生成器A和判别器A）实现从灰度图像到彩色图像的翻译过程,另一个子网络B（包含生成器B和判别器B）反转该过程,即生成器B对称地使用生成器A的最终输出图像作为输入,用来重建原始的人脸灰度图像.其中,网络中的循环损失进行图像重建,而生成损失和对抗损失用来保证生成的图像更加接近真实图像.实验结果表明,这种结构设计不仅能实现自然逼真的人脸图像彩色化,还能同时保证人脸的身份属性不变.      

空间l2(Z×Z)上正交小波基的频域特征刻画与算法

小波基的频域特征刻画有助于小波基的构造。首先给出了函数空间l²(Z×Z)上正交小波基的频域特征刻画，再根据正交小波基的频域特征刻画，可以容易验证：一维空间的两个正交小波基的乘积是二维空间的正交小波基。最后还给出了完美重构条件以及快速分解重构算法。     

Molecular phylogenetics of Gymnocypris （Teleostei：Cyprinidae） in Lake Qinghai and adjacent drainages

149 complete mitochondrial DNA (mtDNA) cytochrome b (Cyt b) genes (1140 bp) of Gymnocypris przewaiskii, Gymnocypris eckloni and Gymnocypris scolistomus from the Lake Qinghai, Yellow River and Qaidam Basin were sequenced and analyzed. Consistent dendrogram indicated that the samples collected from the same species do not constitute a separate monophyletic group and all the samples were grouped into three highly divergent lineages (A, B and C). Among them, Lineage A contained all samples of G.przewaiskii from the Lake Qinghai and partial samples of the G. eckloni from the Yellow River. Lineage B contained the remaining samples of G. eckloni from the Yellow River.Lineage C was composed of a monophyletic group by G. eckloni from the Qaidam Basin. Analysis of molecular variance (AMOVA) indicated that most of genetic variations were detected within these three mtDNA lineages (93.12%), suggesting that there are three different lineages of Gymnocypris in this region. Our Cyt b sequence data showed that G.przewaiskii was not a polytypic species, and G. scolistomus was neither an independent species nor a subspecies of G.eckloni. The divergent mtDNA lineages of G. eckloni from theYellow River suggested that gene flow between the different populations was restricted to a certain extent by several gorges on the upper reach of the Yellow River. Lineage B of G. eckloni might be the genetic effect from the ancestor which was incorporated with the endemic schizothoracinefishes when the headward erosion of the Yellow Riverreached to its current headwaters of late. The G. eckloni from Basin Qaldam was a monophyletic group (lineage C) and Fst values within G. eckloni from the Yellow River were higher than 0.98, suggesting that the gene flow has been interrupted for a long time and the G. eckloni from Basin Qaidam might have been evolved into different species by ecology segregation. The correlation between the rakers number of Gymnocypris and population genetic variation was not significant.All Gymnocypris populations exhibited a low nucleotide diversity (n=0.00096-0.00485). Therefore the Gymnocypris populations from Basin Qaidam could have experienced severe bottleneck effect in history. Our result suggested Gymnocypris populations of Basin Qaidam should give a high priority in conservation programs.     

Mutagenesis of Ser24 of Cytochrome b559 α Subunit Affects PSII Acitivies in Chlamydomonas reinhardtii

In order to study the functions of cytochrome b559 (Cyt b559) in photosystem two (PSII) activity, mutant S24F of Chlamydomonas reinhardtii was constructed using site directed mutagenesis, in which Serine24 (Ser24) locating downstream of Histidine23 (His23) in α subunit of Cyt b559 was replaced by Phenylalanine (Phe). Physiological and biochemical analysis showed that mutant S24F could be grown photoautotrophically or photoheterotrophically. However, their growth rate was slower either on HSM or TAP medium than that of the control; Analysis of PSII activity revealed that its oxygen evolution was about 71% of wild type (WT); The Photochemical efficiency of PSII (Fv/Fm) of S24F was reduced 0.23 compared with WT; S24F was more sensitive to strong light irradiance than the wild type; Furthermore, SDS-PAGE and Western-blotting analysis indicated that the expression levels of α subunit of Cyt b559, LHCII and PsbO of S24F were a little less than those of the wild type. Overall, these data suggests that Ser24 plays a significant role in making Cyt b559 structure maintain PSII complex activity of oxygen evolution although it is not directly bound to heme group.     

Adrenomedullin inhibits the endothelin production induced by urotensin II in rat vascular smooth muscle cells

The aim of this study was to observe the effects of adrenomedullin (ADM) on endothelin (ET) production induced by urotensin Ⅱ (UⅡ) in rat vascular smooth muscle cells (VSMCs). Cultured VSMCs which were incubated with UⅡ (10^-8 mol/L) and with various concentrations of ADM were used to measure the VSMCs ³H-TdR incorpora- tion, the activity of extracellular signal-regulated kinase (ERK), the amount of ET mRNA and ET production in VSMCs. In this work we found that incubation with UⅡ(10^-8 mol/L) increased obviously the amount of ET mRNA in VSMCs and ET production in medium, however, coincubation with ADM (10^-10—10^-8 mol/L) and UⅡ(10^-8 mol/L) reduced the amount of ET mRNA by 15%, 24% and 45% (P< 0.01) respectively, compared with UⅡ alone. The content of ET in medium was 14.13, 11.38 and 11.00 pg/mL. ADM alone (10^-8 mol/L) had no effect on ET production in VSMCs. UⅡ (10^-8 mol/L) promoted the 3H-TdR incorpo- ration and activity of ERK in VSMCs. ADM inhibited VSMCs 3H-TdR incorporation and activation of ERK in a concentration-dependent manner. Compared with UⅡ group, after coincubation with ADM (10^-10—10^-8 mol/L) and UⅡ (10^-8 mol/L) the VSMCs 3H-TdR incorporation was decreased by 7% (P > 0.05), 32% (P < 0.05) and 41% (P < 0.01), respectively, and the activity of ERK was decreased by 24% (P > 0.05), 32% (P < 0.05) and 36% (P < 0.05), re- spectively, in a concentration-dependent manner. The results show that in cultured VSMCs ADM inhibits ET mRNA expression, ET production and proliferation stimulated by UⅡ, and that inhibitory effect of ADM on UⅡ bioaction could be mediated through inhibiting MAPK pathway.     

加权Hardy空间上复合算子的伴随算子的循环性质

讨论加权Hardy空间上复合算子的伴随算子的循环性质,给出cv(Cφ*)在H2(β)中稠密的2个充分条件及Cφ*在H2(β)中循环的1个充分条件.