Phylogenetic Analysis of Field Isolates of Feline Calcivirus (FCV) in Japan by Sequencing Part of Its Capsid Gene

The molecular epidemiology of the infectious disease caused by feline calcivirus (FCV) in Japan was investigated by analysing the phylogenetic relationship among 21 Japanese field isolates, including the F4 strain, and 30 global isolates. Parts of the capsid gene (B–F) of the isolates were amplified by RT-PCR, and the amino acid sequences were compared with those from the global isolates. Thirty-seven and 14 out of a total of 51 isolates were clustered into two distinct genogroups, I and II respectively, by UPGMA and NJ analysis. Seven of the 21 Japanese isolates (33%) fell into group I together with 30 global isolates, while the other 14 Japanese isolates (67%) belonged to group II. The bootstrap repetition analysis of groups I and II formed by the NJ method gave a value of 99.0%. The 14 latter Japanese isolates were clearly separated from the isolates in group I, and they were different from any previously known FCV, forming a new genogroup, which implies that this lineage has been confined to Japan. Comparing the amino acid sequences shared by groups I and II, the amino acid at position 377 in B region was asparagine (Asn or Asp (NH₂)) in group I, while it was lysine (Lys) in all the strains in group II. Similarly, the amino acid at position 539 in the F region was alanine (Ala) or proline (Pro) in group I, while it was valine (Val) in group II; glycine (Gly) at position 557 in group I was serine (Ser) in Group II; and phenylalanine (Phe) or leucine (Leu) at position 566 in genogroup I was tyrosine (Tyr) in group II.     

一类分式序列封闭形和式

利用反正切函数关系arctan F（n）-arctan F（n＋1）=arctan （F（n）-F（n＋1））/（1＋F（n）F（n＋1））得到一类反正切序列封闭形和式,利用微分法得到一类分式序列封闭形和式,并给出反正切级数与分式级数恒等式.     

Outbreak of Philophthalmus gralli in four greater rheas (Rhea americana)

Using slit‐lamp biomicroscopy, conjunctival biopsy, and morphological identification, a flock of four Greater rheas (Rhea americana) in Arizona were diagnosed with conjunctivitis secondary to Philophthalmus gralli (P. gralli) infection. Aquatic snails from the exhibit’s water source were identified as Melanoides tuberculatus, a known vector for P. gralli. Comparison of partial sequences of DNA regions from P. gralli adults removed from the rheas and metacercariae from the aquatic snails demonstrated a 100% match, confirming the source of infection. The flock was divided into two treatment groups: the most severely affected rheas received both manual removal of trematodes and praziquantel 1% ointment OU q12 h and the least severely affected rheas were only given praziquantel 1% ointment OU q12 h. The rheas were permanently relocated away from the infected water source and aquatic snails. Initial resolution was seen at 17 weeks in the most severely affected rhea, which had 675 adult P. gralli removed and topical praziquantel. The two rheas that only received topical praziquantel showed resolution within 3 and 15 weeks. Current recommendations for treating P. gralli include: manual removal of trematodes, topical praziquantel 1% ointment, and relocation away from infected water sources and aquatic snails.     

禾本科8种牧草DNA条形码通用序列筛选

DNA条形码技术是利用DNA保守片段对物种进行快速准确鉴定的新兴技术。本研究根据GenBank中禾本科牧草matK和rbcL基因的核苷酸序列,设计4对通用引物,建立并优化了针对禾本科7个主要牧草属8种牧草11个样品[高丹草(Sorghum bicolor×S.sudanense)、玉米(Zea mays)、针茅(Stipa capillata)、"贝克"多年生黑麦草(Lolium perenne‘Plxie)、"凯帝莎"多年生黑麦草(L.perenne‘Caddieshack’)、甘肃羊茅(Festuca kansuensis)、"百琪"紫羊茅(F.rubra‘Bargena’)、"梦神"紫羊茅(F.rubra‘Maxima’)、"百胜"草地早熟禾(Poa pratensis‘Barvictor’)、"钻石"草地早熟禾(P.pratensis‘Diamond’)和芨芨草(Achnatherum splendens)]的目的片段的扩增条件。对扩增产物进行测序和分析,经分别比对,筛选出8种牧草的4个标记位点5’端和3’端保守序列。对各标记位点保守区内的核苷酸进行单核苷酸多态性(SNPs)的单倍型分析。结果表明,matK1、matK2、matK3分别有6个单倍型(H1~A、H1~B、H1~C、H1~D、H1~E和H1~F)、7个单倍型(H2~A、H2~B、H2~C、H2~D、H2~E、H2~F和H2~G)和3个单倍型(H3~A、H3~B和H3~C),rbcL基因有5个单倍型(H4~A、H4~B、H4~C、H4~D和H4~E)。根据matK(matK1、matK2、matK3)和rbcL基因筛选的4个标记位点为8种牧草建立了相对应的特异DNA识别码。本研究可为混合禾本科牧草饲料中的高粱属、玉蜀黍属、芨芨草属、针茅属、黑麦草属、羊茅属和早熟禾属的8种牧草准确识别提供分子水平上的科学依据。     

Identification of a new family of tandem repeats in Triticeae genomes

A new family of cereal tandem repeats was isolated, characterised and designated as spelt-1. The family of repeats comprises about 2% of the Aegilops speltoides genome; however, its content differs considerably in the genomes of various Triticeae species. Copy number of the constituent sequence, relative to Ae. speltoides, proved to be 40-60 times reduced in the genomes of tetraploid wheats, 400-fold reduced in the genome of Triticum monococcum, and 1200-2400 times in the genomes of the other 19 Triticeae species studied. Drastic difference in the copy number and homology extent of the spelt-1 family sequences between Ae. speltoides and other diploid species allows the utilisation of these sequences as species-specific telomeric markers for Ae. speltoides, provided stringent hybridisation conditions apply. RFLP (restriction fragment length polymorphisms) analysis of spelt-1 reveals polymorphism between the above species. This study of spelt-1 organisation in different Triticum species provided further substantiation of the polyphyletic origin of the B genome of polyploid wheat. This revised version was published online in August 2006 with corrections to the Cover Date.     

牙鲆碱性磷酸酶基因调控序列的功能分析及甲状腺激素对其调节作用

为分析牙鲆碱性磷酸酶基因5'调控区和第一内含子调控序列的功能,本研究运用DNA重组技术将PCR扩增得到的牙鲆碱性磷酸酶基因的5′调控区序列、第一内含子序列及5′调控区和第一内含子串联序列分别插入启动子缺失的增强型绿色荧光蛋白表达载体pAcGFP1-1中,得到pAcGFP1-5′ALP、pAcGFP1-Intron1和pAcGFP1-AI报告基因表达载体。通过脂质体介导重组质粒转染中国仓鼠卵巢细胞(CHO)并检测荧光信号。转染后细胞状态良好,24 h后发现,在倒置荧光显微镜下均可看到不同强度的绿色荧光信号,且荧光信号随时间的延长而增强,其中转染pAcGFP1-AI后的荧光信号强度最强。以上结果表明,牙鲆碱性磷酸酶基因5′调控区和第一个内含子序列均具有启动子活性,且第一内含子与5′调控区存在协同效应,两者协同能增强基因的表达。为了解甲状腺激素T3对调控序列启动子活性的调节作用,用不同浓度的T3处理转染的CHO细胞,24 h后发现加入50、75和100 nM的T3都增强了绿色荧光蛋白的表达,而且信号强度随T3浓度的增加而增强。这表明甲状腺激素T3对牙鲆碱性磷酸酶基因调控序列的启动子活性有一定的增强作用。本研究为深入阐明牙鲆碱性磷酸酶基因转录表达的分子机制提供了实验依据。     

Biochemical comparison of lectins among three different color strains of the red alga Kappaphycus?alvarezii

The red alga Kappaphycus alvarezii is economically important as an edible species and as a source of carrageenan, and has been extensively cultivated in many tropical countries. For this species, different color strains, which differ from each another in growth rate and carrageenan content, have been reported for decades. In this study, lectins from brown, red, and green strains of K. alvarezii cultivated in Vietnam were isolated and characterized for evaluation of their biochemical properties and contents. The results showed that each color strain contained in common the three lectins, named KAA-1, KAA-2, and KAA-3, which shared the hapten-inhibition profile of hemagglutination, 20 N-terminal amino acid sequence, and equivalent molecular mass within a range of 28,016 ± 1.2 to 28,021 ± 1.8 Da, but differed in their yields, with the highest yield of KAA-2. These properties of the three isolectins were also comparable among the three different color strains. However, the sum of the yields of the three isolectins decreased in the order: red (21.4 mg) to green (15.9 mg) to brown strains (15.1 mg), from 500 g fresh alga. Thus, this algal species can be a good source of useful lectins, irrespective of color strain.     

基于细胞色素b基因序列的鲌亚科鱼类系统发育研究

采用特异性引物PCR扩增获得了鲌亚科7属10种鱼类细胞色素b基因的全长序列,初步构建了鲌亚科鱼类的系统发育关系。NJ树和MP树一致表明,鲌亚科鱼类种内所有个体均单独聚群,支持率高;同一属内,除蒙古鲌(Culter mogolicus mongolicus)未能与同属的翘嘴鲌(Culter alburnus)、青梢鲌(Culter dabryi dabryi)单独聚为一支外,团头鲂(Megalobrama amblycephala)与三角鲂(Megalobrama terminal)、翘嘴鲌与青梢鲌均为姐妹种。NJ、MP树显示鲌亚科鱼类属间关系的支持率都较低,鲂属(Megalobrama)与鳊属(Parabramis)间、鲌属(Culter)与原鲌属(Cultrichthys)间、细鳊属(Rasborinus)与半属(Hemiculterella)、属(Hemiculter)间有较近的亲缘关系,在系统演化关系上,细鳊属、半属、依氏鱼属(Ishikauia)较为原始,鲌属与原鲌属、鲂属与鳊属互为姐妹群,为较特化群。     

海南4个地区暹罗炭疽菌群体遗传结构分析

对分离自海南4个地区13种寄主植物上暹罗炭疽菌的ITS-CAL-GAPDH 3基因序列进行群体遗传结构分析。95条暹罗炭疽菌多基因序列可定义为16个单倍型,其中,单倍型H5为主要单倍型,分布于所有寄主植物。AMOVA分析显示,遗传变异主要发生在种群内;病菌未经历过大规模的种群扩张过程。研究结果表明,暹罗炭疽菌种群具有丰富的遗传多样性。     

鸡卵清蛋白基因调控序列的克隆与载体构建

卵清蛋白（ovalbumin）基因在鸡基因组中只有一对等位基因,却能每天合成分泌多达2 g的蛋白,占据卵白蛋白质的50%以上,是外源基因载体表达调控构件的首选。该研究从输卵管特异性启动子方面着手,通过对卵清蛋白基因启动子的筛选优化,找出启动子增强因子的位置以及组织特异性区域。将卵清蛋白基因上游-922～-2 073和-2 801～-3 100区域平均分成12个长度约为150 bp的序列,分别插入到-921～+38序列的上游,成功构建12个系列表达载体,为进一步筛选短缩版优化启动子提供材料;卵清蛋白基因第一内含子区域截断成300 bp左右的迷你内含子序列,成功构建8个迷你内含子系列载体,为筛选优化的迷你内含子提供必要的材料;成功分离鸡输卵管上皮细胞并优化电转染条件,通过荧光素酶活性检测初步筛选出具有最强活性重组质粒pGL4 UP 1412和内含子重组质粒pGL4 mini intron 3,同时推断出若干包含增强子序列区域。