Genetic diversity of Chinese domestic goat based on the mitochondrial DNA sequence variation

The aim of this study was to characterize the genetic diversity of domestic goat in China. For this purpose, we determined the sequence of the mitochondrial DNA (mtDNA) control region in 72 individuals of the Yangtze River delta white goat, and reanalysed

Identification of differential transcript profiles between mutual crossbred embryos of zebrafish (Danio rerio) and Chinese rare minnow (Gobiocypris rarus) by cDNA-AFLP

The crosstalk between naive nucleus and maternal factors deposited in egg cytoplasm before zygotic genome activation is crucial for early development. In this study, we utilized two laboratory fishes, zebrafish (Danio rerio) and Chinese rare minnow and Chinese rare minnow (Gobiocypris rarus), to obtain mutual crossbred embroys and examine the interaction between nucleus and egg cytoplasm from different species. Although these two types of crossbred embryos originated from common nuclei, various developmental capacities were gained due to different origins of the egg cytoplasm. Using cDNA amplified fragment length polymorphism (cDNA-AFLP), We Compared transcript profiles between the mutual crossbred embryos at two developmental stages (50%- and 90%-epiholy). Three thousand cDNA fragments were generated in four cDNA pools with 64 primer combinations. All differently displayed transcript-derived fragments (TDFs) were screened by (lot blot hybridization, and the selected sequences were further analyzed by semi-quantitative RT-PCR and quantitative real-time RT-PCR. Compared with ZR embryos, 12 genes were up-regulated and 12 were down-regulated in RZ embryos. The gene fragments were sequenced and subjected to BLASTN analysis. The sequences encoded various proteins which functioned at various levels of proliferation, growth, and development. One gene (ZR6), dramatically down-regulated in RZ embryos, was chosen for loss-of-function study; the knockdown of ZR6 gave rise to the phenotype resembling that of RZ embryos. (c) 2008 Elsevier Inc. All rights reserved.

剪切流动法研究表面固定化配基与目标分子的相互作用力

基于剪切流动腔技术,以微球作为受力载体,设计了一套可用于研究表面固定化配基与目标分子特异性相互作用力的实验和分析方法,并以人免疫球蛋白G (human IgG)和羊抗人免疫球蛋白G(goat anti-human IgG)分别作为模型配基和模型目标分子进行了研究.基于平面Poiseuille层流模型设计了流场参数,以数值计算结果验证了设计的合理性.使用牛血清白蛋白(BSA)作为非特异性对照,判断微球与基片表面的结合力来自配基和目标分子的生物特异性相互作用,并由进一步的目标分子灭活对比实验确认了这一结论.实验观察到微球与基片表面的结合力受到配基面密度的影响,说明发生结合的是多对而非单对蛋白质分子.将95%的微球被剥离时对应的壁面剪切率设定为临界剪切率,由大量实验结果拟合得到了临界剪切率与配基面密度间的定量关系.在受力分析模型中,考虑到多分子的结合,以及分子键位置不同造成的力臂长度的差异,最终计算得到单对配基与目标分子的平均结合力约为342pN.

直墙前破波波浪力──对修正过的Gota方法的探讨

对被重新修正后的Goat在墙波压力计算方法进行了分析，主要是对其无因次波浪力随基床肩宽和相对波高的变化进行分析，结果发现，Goat法计算的无因次波浪力随基床肩宽的变化，有一最不利肩宽，此不利肩宽与相对波高无关；在同一水深条件下，Goat法计算的无因次波浪力随相对波高的增大而增大。此两点与试验数据不符。

Study of Interaction Force between Antigen and Antibody Using Flow Chamber Method

A parallel plate flow chamber was used to study the interaction force between human IgG (immobilized on a chip surface as ligand) and goat anti-human IgG (immobilized on microspheres surface as receptor). First, it was demonstrated that the binding force between the microspheres and the chip surface came from the bio-specific interaction between the antigen and the antibody. Secondly, it was obtained that the critical shear rate to detach microspheres from the chip surface increases with the ligand surface concentration. Finally, two models to estimate the antigen-antibody bond strength considering bonds' positions were proposed and analyzed.

Phylogenomic study of the subfamily Caprinae by cross-species chromosome painting with Chinese muntjac paints

Chromosomal homologies have been established between the Chinese muntjac (Muntiacus reevesi, MRE, 2n = 46) and five ovine species: wild goat (Capra aegagrus, CAE, 2n = 60), argall (Ovis ammon, OAM, 2n = 56), snow sheep (Ovis nivicola, ONI, 2n = 52), red goral (Naemorhedus cranbrooki, NCR, 2n = 56) and Sumatra serow (Capricornis sumatraensis, CSU, 2n = 48) by chromosome painting with a set of chromosome-specific probes of the Chinese muntjac. In total, twenty-two Chinese muntjac autosomal painting probes detected thirty-five homologous segments in the genome of each species. The chromosome X probe hybridized to the whole X chromosomes of all ovine species while the chromosome Y probe gave no signal. Our results demonstrate that almost all homologous segments defined by comparative painting show a high degree of conservation in G-banding patterns and that each speciation event is accompanied by specific chromosomal rearrangements. The combined analysis of our results and previous cytogenetic and molecular systematic results enables us to map the chromosomal rearrangements onto a phylogenetic tree, thus providing new insights into the karyotypic evolution of these species.

山羊mtDNA 多态性及其起源分化研究

　本文用14 种识别6 碱基的限制性内切酶Apa Ⅰ、BamH Ⅰ、Bgl Ⅰ、Bgl Ⅱ、Dra Ⅰ、 EcoR Ⅰ、EcoR Ⅴ、Kpn Ⅰ、Pvu Ⅱ、Pst Ⅰ、Sac Ⅰ、Sal Ⅰ、Sma Ⅰ和Xho Ⅰ研究了来自我国12 个省 和自治区共计18 个地方山羊品种218 个个体mtDNA 的RFL P ,并运用Nei 氏公式计算了各限制 性类型间的遗传距离P 和群体遗传多态度π值。结果表明,在研究的所有个体中共检测到41 个 酶切位点,18 种限制性态型,其中BamH Ⅰ、Bgl Ⅱ、Dra Ⅰ、EcoR Ⅰ和Sal Ⅰ共5 种酶表现出多态。 18 种限制性态型可归结为6 种基因单倍型,单倍型Ⅰ(BamH Ⅰ2B、Bgl Ⅱ2A、Dra Ⅰ2A、EcoR Ⅰ2A 和Sal Ⅰ2A) 和单倍型Ⅱ(BamH Ⅰ2B、Bgl Ⅱ2A、Dra Ⅰ2A、EcoR Ⅰ2A 和Sal Ⅰ2B) 为两种基本单倍 型,研究结果提示我国地方山羊品种起源于两种不同的母系祖先。各限制性类型间的平均遗传距 离为0100436 ,整个群体的平均遗传多态度π值为010487 % ,表明我国地方山羊品种mtDNA 遗传 多样性比较贫乏,分化程度较低。

黔东南小香羊与贵州原有其它山羊品种的线粒体DNA 多态性比较

用B am H É、B g lÉ、B g lÊ、D raÉ、E coR É、E coR Í、H indË、Kp nÉ、 P stÉ、P vuÊ、S acÉ、S a lÉ、Sm aÉ、S tuÉ、X hoÉ 15 种识别6 碱基的限制性内 切酶对黔东南小香羊和贵州原有3 个地方山羊品种的93 只个体的m tDNA 进行分 析表明, B am H É、H indË 和S a lÉ 3 种酶表现多态性; 共检出18 种限制性多态型, 归纳得到3 种单倍型, 以单倍型É 和Ê 为基本单倍型。根据此2 种基本单倍型在所 比较各品种中的不同分布比例, 以及遗传距离分析和品种间的聚类关系, 表明黔东 南小香羊的群体遗传构成与贵州省原有其它3 个山羊品种不同, 从而为进一步确认 其为一独立的品种提供了必要的分子生物学依据。

Mitochondrial diversity and phylogeographic structure of Chinese domestic goats

China has numerous native domestic goat breeds, but so far there has been no extensive study on genetic diversity, population demographic history, and origin of Chinese goats. Here, we examined the genetic diversity and phylogeographic structure of Chinese domestic goats by determining a 481-bp fragment of the first hypervariable region of mitochondrial DNA (mtDNA) control region from 368 individuals representing 18 indigenous breeds. Phylogenetic analyses revealed that there were four mtDNA lineages (A-D) identified in Chinese goats, in which lineage A was predominant, lineage B was moderate, and lineages C and D were at low frequency. These results further support the multiple maternal origins of domestic goats. The pattern of genetic variation in goat mtDNA sequences indicated that the two larger lineages A and B had undergone population expansion events. In a combined analysis of previously reported sequences and our sequences belonging to lineage B, we detected two subclades, in which one was unique to eastern Asia and another was shared between eastern and southern Asia. A larger genetic variation in eastern Asia than southern Asia and the pattern of phylogeographic variation in lineage B suggest that at least one subclade of lineage B originated from eastern Asia. There was no significant geographical structuring in Chinese goat populations, which suggested that there existed strong gene flow among goat populations caused by extensive transportation of goats in history. (c) 2005 Elsevier Inc. All rights reserved.

云南保种山羊线粒体DNA限制性酶切研究

　采用碱变性法,提取来自云南省不同地区4个保种山羊的13个个体的线粒体DNA(mtDNA),并用ApaⅠ,AvaⅠ,BamHⅠ,BclⅠ,BcIⅠ，BglⅡ,ClaⅠ,DraⅠ, EcoRⅠ,EcoRⅤ,HaeⅠ,HindⅢ,KpnⅠ,PstⅠ,PvuⅡ,SacⅠ,SalⅠ,SmaⅠ,StuⅠ和XhoⅠ等20种限制性内切酶进行酶切分析。结果发现它们的线粒体DNA的分子量大 小约为15.8Kb；不同限制性内切酶的酶切位点分别为：DraⅠ有7个酶切位点，AvaⅢ有6个酶切位点，EcoRⅤ和StuⅠ共有5个酶切位点，HindⅡ和HaeⅡ有4个酶 切位点，BamHⅠ,BglⅡ,PstⅠ和PvuⅡ有3个酶切位点，ApaⅠ，ClaⅠ有两个酶切位点，其余有1个酶切位点。各保种山羊间未发现变异，说明云南的4个保种山 羊极可能来自于共同的母性祖先。

不同世代改良型路南奶山羊血液免疫学参数的比较研究

本文测定了“改良型路南奶山羊”的亲本及改良不同世代 (F1,F2 ,F3,F4 )母羊的外周血T淋巴细胞转化率、ANAE阳性率、B淋巴细胞ZYC花环率、红细胞IC花环率、C3b花环率以及血清γ -球蛋白等免疫学参数 ,比较研究其随着世代增递的变异情况。结果表明 :从亲代到F4代随着改良世代的递增 ,T淋巴细胞转化率、ANAE阳性率、红细胞C3b花环率和IC花环率和血清γ -球蛋白含量均呈F1>F2 >P >F3>F4变化 ,ZYC花环率呈F1>P >F2 >F3>F4变化。回归分析的结果表明 :从亲代到改良四代 ,上述各项免疫学参数均呈显著负相关 ,趋势线为下降趋势。即随着世代的递增有逐代递减的趋势。结果提示 :用西农莎能羊改良路南圭山羊 ,其改良一代和二代的上述各项血液免疫学参数均略高于路南圭山羊 ,以后随着世代的递增有逐代递减的趋势。预示着随着世代的递增 ,改良型路南奶山羊的体质状况 ,免疫功能和抗病能力可能会出现下降趋势。

The first non-mammalian CXCR3 in a teleost fish: Gene and expression in blood cells and central nervous system in the grass carp (Ctenopharyngodon idella)

A novel fish chemokine receptor gene, chemokine (C-X-C motif) receptor 3 (CXCR3)-like was isolated from the grass carp Ctenopharyngodon idella , with its full-length genomic sequence. The cDNA of grass carp CXCR3-like (gcCXCR3-like) consists of 1261 bp with a 49bp 5'-UTR and a 189 bp 3'-UTR. An open reading frame of 1023 bp encodes a 341-amino acid peptide, with seven transmembrane helices. The deduced amino acid sequence showed the same sequence identities (37.8%) with its counterparts in goat and human. The gcCXCR3-like gene consists of two exons, with one intervening intron, spaced over approximately 2 kb of genomic sequence. Phylogenetic analyses clearly demonstrated that the gcCXCR3-like resembles the CXCR3s of other vertebrates. Real-time PCR analysis showed that gcCXCR3-like was expressed in all tested organs except heart and the expression level of gcCXCR3-like was highest in brain. Flow cytometric analyses showed the positive rate of labelled leukocytes from the healthy grass carp was 17.3%, and the labelled leukocytes were divided into three types by cell sorting. Immunohistochemical localization revealed that gcCXCR3-like expressed in whole brain regions including cerebel, diencephalon, medulla oblongata, optic lobe, and rhinencephalon, and that the labelled leukocytes are actually populations of monocyte and/or phagocyte, lymphocyte and the granulocyte. It is considered that fish CXCR expression and their function may need to be investigated in both nervous and immune systems. (c) 2006 Elsevier Ltd. All rights reserved.

Preparation of pH-sensitive polymer by thermal initiation polymerization and its application in fluorescence immunoassay

A fluorescence immunoassay for human IgG (Ag) was developed using a pH-sensitive polymer prepared by thermal initiation or redox initiation polymerization as a carrier. In the competitive immunoassay, appropriate quantity of Ag was immobilized on the polymer and the standard Ag (or sample) solution, and a constant amount of fluorescein isothiocyanate labeled goat anti-human IgG antibody (Ab-FITC) was added. Immobilized Ag and the standard (or sample) Ag competed for binding to the Ab-FITC in 37 C in homogeneous format. After changing the pH to separate the polymer-immune complex precipitate, it was re-dissolved and determined by fluorescence method. The results showed that the immobilization efficiency, immunological reaction activities of immobilized Au and phase transition pH range were improved as Ag was immobilized by thermal initiation instead of redox initiation polymerization. Under optimum conditions, the calibration graphs for the Ag in both methods, thermal initiation and redox initiation, were linear over the concentration range of 0.0-1000 ng mL(-1), with detection limits 8 (thermal initiation) and 12 ng mL(1) (redox initiation), respectively. Moreover, some pH-sensitive polymer prepared only in organic solvent or under high temperature could also be used as an immunoreaction carrier by thermal initiation polymerization. Thermal initiation polymerization was a better immobilization mode. (C) 2004 Elsevier B.V. All rights reserved.

Electrochemical Metalloimmunoassay Based on Enlargement of Gold Nanoparticle Label Treated with Ag Enhancement System

A novel sensitive electrochemical immunoassay with colloidal gold as the antibody labeling tag and subsequent signal amplification by silver enhancement is described. Colloidal gold was treated by a light-sensitive silver enhancement system which made silver deposit on the surface of colloidal gold(form Au/Ag core-shell structure), followed by the release of the metallic silver atoms anchored on the antibody by oxidative dissolution of them in an acidic solution and the indirect determination of the dissolved Ag+ ions by anodic stripping voltammetry(ASV) at a carbon fiber microelectrode. The electrochemical signal is directly proportional to the amount of analyte(goat IgG) in the standard or a sample. The method was evaluated by means of a noncompetitive heterogeneous immunoassay of immunoglobulin G(IgG) with a concentration as low as 0.2 ng/ mL. The high performance of the method is related to the sensitive ASV determination of silver(I) at a carbon fiber microelectrode and to the release of a large number of Ag+ ions from each silver shell anchored on the analyte(goat IgG).

Experimental evaluation of heterobeltiosis and heterosis between two populations of Pacific abalone Haliotis discus hannai Ino

A2 x 2 factorial cross between two populations of Pacific abalone Haliotis discus hannai Ino, collected separately from Dalian (D) in China and Miyagi (M) in Japan, was conducted to compare performances in fertilization rate, hatching rate, metamorphosis rate and growth at days 20, 43, 160 and 330 between purebreds (DD and MM) and crossbreds (DM and MD) and investigate the magnitude of heterobeltiosis (better parent) and heterosis (mid-parent). Heterobeltiosis and heterosis for all the traits analyzed were evidently different between crossbreds DM and MD. Heterobeltiosis in the crossbred DM varied among traits, with values of 2.5% for the fertilization rate, 2.2% for the hatching rate, -1.9% for the metamorphosis rate and 7.4% for the growth at the (lay 330. The crossbred DM displayed positive heterotic values for fertilization rate (5.4%), hatching rate (7.4%), metamorphosis rate (7.6%) and growth (12.0%) at the day 330. However, both heterobeltiosis and heterosis for all the traits in the crossbred MD were negative except those for the growth at days 20 and 43. The results indicate the importance of selecting superior hybrid varieties if the exploitation of hybrid vigor is considered in the Pacific abalone breeding program.