封闭群比格犬微卫星的筛选及初步应用

摘要:目的 筛选可用于比格犬遗传检测的微卫星位点,并对小型比格犬群体进行了遗传结构分析。 方法 选取来自文献的微卫星位点 120 个,包括比格犬位点 23 个和其他犬类遗传检测的位点 97 个。 用 DNA 池对这些位点进行 PCR 条件优化后,挑选扩增效果优良的位点对常用比格犬群体进行荧光标记 STR 扫描和群体遗传结构分析,依据期望杂合度( He)和香农指数挑选符合要求微卫星位点计算群体遗传参数。 结果 在候选的 120 个位点中共有73 个位点扩增效果优良,STR 扫描后选用 49 个位点进行遗传分析,结果显示该群体有效等位基因数、期望杂合度、平均杂合度、香农指数、多态性信息含量分别为 4. 9230、0. 7574、0. 7375、1. 6625、0. 7038,证明该群体遗传多态信息丰富,群体遗传多样性高。 结论 所选 49 个微卫星位点可用于比格犬遗传检测,这些位点还需用更多群体进行验证和优化。     

基于微卫星标记的中华蜜蜂5个自然群体遗传多样性分析

【目的】分析中国四川阿坝、黄土高原、海南岛、浙闽丘陵、滇南地区等地的中华蜜蜂(Apis cerana cerana)自然群体遗传多样性。【方法】提取样品基因组DNA后进行PCR扩增,并用聚丙烯酰胺凝胶电泳检测。通过计算15对微卫星标记在5个中华蜜蜂自然群体中的等位基因数、多态信息含量、期望杂合度、观测杂合度、遗传距离、遗传相似系数和遗传分化系数,进而评估各中华蜜蜂种群遗传多样性和各种群间遗传分化。【结果】15个微卫星位点共检测到64个等位基因,每个位点的等位基因数平均为4.266 7个,单个位点的等位基因数为2～6个。各群体平均等位基因数为2.6～3.066 7个,平均观测杂合度为0.550 3～0.664 3,平均期望杂合度为0.524 4～0.566 7,平均多态信息量为0.410 3～0.492 7。四川阿坝群体与黄土高原、海南岛、浙闽丘陵、滇南地区等地群体在Acc001,Acc096,Acc131等3个位点上存在明显分化;黄土高原群体与海南岛、浙闽丘陵、滇南地区等地群体在Acc114,Acc121,Acc132等3个位点上表现出一定程度的遗传分化;海南岛群体与浙闽丘陵、滇南地区两地群体在Acc001,Acc002,Acc004,Acc131,Acc132等5个位点上均表现出较强的遗传分化;浙闽丘陵与滇南地区群体间在Acc002,Acc121,Acc131等3个位点上遗传分化极明显。5个中华蜜蜂自然群体之间的遗传距离在0.160 1～0.491 0之间。【结论】5个中华蜜蜂群体遗传多样性丰富,群体间遗传分化明显。     

舟山附近海域3个大黄鱼养殖群体遗传多样性的微卫星分析

利用8个微卫星多态标记分析舟山附近海域3个大黄鱼养殖群体共90个体的遗传多样性。结果显示,8个微卫星位点共检测到等位基因64个,各位点等位基因数(A)范围为5~10个,观测杂合度(Ho)的平均值为0.9111,期望杂合度(He)的平均值为0.8108,多态信息含量(PIC)平均值为0.7800,表明所选择的8个微卫星位点均表现出较高的多态性。群体的遗传多样性分析结果显示,3个群体的平均观测杂合度和平均期望杂合度分别为0.891 7、0.900 0、0.941 7和0.738 4、0.743 2、0.821 3,Shannon多样性指数分别为1.467 0、1.449 0、1.800 9,表明3个群体的遗传多样性处于较高水平。HardyWeinberg平衡检验发现只有LYC0015和LYC0009位点处于平衡状态(PHW0.05),其余6个位点都不同程度地偏离了平衡。3个群体的聚类分析表明,岱山和朱家尖西岙网箱2个养殖群体亲缘关系较近,而与舟山市水产所耐低温F2代群体亲缘关系较远。     

应用T载体连接PCR技术分离黄嘴白鹭微卫星位点

传统的微卫星分离策略对于低丰度的物种如鸟类而言,通常是低效的.探讨了一种改良的T载体PCR序列获取法用于黄嘴白鹭(Egretta eulophotes)微卫星DNA的分离,称为T载体连接PCR(T-vector-ligation PCR,TVL-PCR).在获取第一侧微卫星序列的71个阳性克隆中,59个克隆含有重复序列,设计了48对位点特异巢式引物.经过两轮TVL-PCR后,31个位点的扩增产物为预期大小,经测序后获得21个完整的微卫星位点.除去侧翼序列过短和小卫星位点外,设计16个微卫星位点进行多态信息检测,其中7个位点显示出多态性,平均等位基因数为2～20,观察杂合度(HO)和期望杂合度(HE)分别为0.375～0.958和0.477～0.956,所有位点都符合哈迪-温伯格平衡(HWE)和连锁平衡.说明TVL-PCR技术对低丰度微卫星物种的位点分离具有一定的应用价值.     

基于转录组测序的波纹巴非蛤微卫星标记研究

应用MISA软件对波纹巴非蛤转录组测序数据进行分析,共获得7378个微卫星标记,从中选取95个位点设计引物,并在一个野生波纹巴非蛤群体中进行分析验证,结果显示:95个位点中能够扩增出清晰条带的位点有30个,其中15个位点表现为单态,15个位点呈现多态.使用15个多态位点进行波纹巴非蛤群体的遗传多样性分析,结果表明:这15个位点的等位基因数为2~9,平均等位基因数为4.733,平均有效等位基因数为2.438,平均观测杂合度和平均期望杂合度分别为0.270、0.516,平均多态信息含量为0.457.     

新疆7个绵羊品种MHC区段微卫星遗传多样性的分析

为分析新疆北疆地区主要绵羊品种的遗传多样性和系统发生关系,利用SSR Hunter软件寻找5个微卫星标记,采用PCR扩增,1%琼脂糖凝胶电泳,对新疆7个品种绵羊群体遗传多样性进行了检测分析,统计了各群体的等位基因组成、平均有效等位基因数和平均基因纯合率,利用等位基因频率计算出各群体的平均遗传杂合度、多态信息含量和群体间的遗传距离。结果显示新疆7个绵羊群体共发现28个等位基因,实验证明在5个微卫星位点的平均多态信息含量为0.5569~0.7335,平均杂合度为0.6208~0.7437,有效等位基因数为2.7118~4.4203,均属于高度多态性位点。聚类分析和主成分分析结果与其起源、育成历史及地理分布基本一致。这5个位点作为与生产性能相关的遗传标记较为理想。     

宝石鲈与淡水黑鲷及其子一代微卫星位点分析

使用11对微卫星引物对杂交父本宝石鲈(Scortum barcoo)和母本淡水黑鲷(Hephaestus fuliginosus)及其子一代胜斑(F1)进行分子遗传机制研究,结果表明:(1)在11个微卫星位点中,淡水黑鲷有10个位点得到扩增产物,F1有9个位点得到扩增产物,而宝石鲈只得到6个位点的扩增产物,5个位点能在3个种类中稳定扩增;(2)宝石鲈、淡水黑鲷及其F1这3个群体在这5个位点中的平均等位基因数(Ao)为2.000 0、3.000 0、2.200 0;平均有效等位基因数(Ae)分别为2.000 0、2.719 8、2.133 3;平均观测杂合度(Ho)均为1.000 0;平均期望杂合度(He)分别为0.545 5、0.672 7、0.572 7;平均多态信息含量(PIC)为0.375 0、0.537 6、0.410 9;平均Shannon信息指数(I)分别为0.693 1、1.022 7、0.762 5;(3)F1与宝石鲈和淡水黑鲷的遗传相似性系数分别为0.974 7和0.839 8,遗传距离分别为0.025 6和0.174 6.根据Nei's遗传距离建立的UPGMA聚类图表明杂交F1与2亲本的遗传差异不是对等的,而是偏向父本宝石鲈一方.     

四个近交系斑马鱼微卫星多态性研究

目的研究微卫星DNA多态性在四个近交系实验用斑马鱼遗传监测中的应用。方法选取斑马鱼不同染色体上的3个微卫星位点,应用PCR技术对常用的4种近交系(AB,TU,WIK,LF)斑马鱼进行了微卫星DNA多态性分析。结果3个微卫星DNA具有稳定扩增效果在不同品系之间表现显著多态性。结论运用所筛选的3个位点进行微卫星多态性分析,能够快速、经济地对AB,TU,WIK,LF四种品系斑马鱼进行遗传监测。     

自发性先天性白内障大鼠遗传状况分析

目的 了解自发性先天性白内障(TSC)大鼠的遗传状况，评价遗传质量分析方法。方法 对TSC大鼠、BN大鼠、F344大鼠下颌骨11项变量进行测量，使用SPSS18.0统计软件进行数据分析；选取近交系大鼠常用的6个微卫星位点，通过PCR扩增和琼脂糖电泳鉴定，进行遗传质量检测。结果 TSC大鼠与BN大鼠相比较，下颌骨参数有9个差异有统计学意义(P<0.01);TSC大鼠与F344大鼠相比较，下颌骨参数有5个差异有统计学意义(P<0.01)。TSC大鼠不同个体在6个微卫星位点上的扩增产物均只出现一个条带，4个位点可以将TSC大鼠与BN大鼠区别，4个位点可以将TSC大鼠与F344大鼠区别，1个位点可以将3种近交系大鼠区分。结论 TSC大鼠下颌骨变量与BN大鼠和F344大鼠有明显的差异；TSC大鼠符合近交系要求，包含6个微卫星位点组合，可用于TSC大鼠与常用近交系大鼠品系BN和F344的遗传背景监测。     

果子狸多态性微卫星位点的筛选及特性分析

以采自四川平武县老河沟自然保护区的一份果子狸(Paguma larvata)组织样品为实验材料, 通过FIASCO法构建微卫星文库, 对250个单克隆进行测序分析, 获得147条含微卫星序列的片段, 微卫星单元重复次数大于10的序列共有42条。据此设计引物 42对, 进一步使用21份果子狸样品检测这些引物的扩增能力和扩增序列多态性, 最终得到5个多态性微卫星位点。同时, 对已发表的13个果子狸微卫星位点进行检测, 发现其中5个位点具有多态性。对这10个位点的特性进行分析, 结果显示它们具有较高的多态性(等位基因数为2~11, 观测杂合度为0.286~0.737, 期望杂合度为0.358~0.906); PID和PID-sib值表明, 在约10⁹个没有亲缘关系的个体或10⁴个有亲缘关系的个体中, 可能发现一对基因型相同的个体, 由于果子狸野外局域种群规模远低于这个量级, 这10个微卫星位点可用于果子狸的个体识别。     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

橡胶颗粒沥青混合料低温抗裂性试验研究

以AC-13级配为基础,将橡胶颗粒代替部分集料掺入混合料中,以低温弯曲试验为评价方法对不同橡胶颗粒掺量下沥青混合料的低温抗裂性进行研究,并引入应变能密度值对混合料的低温抗裂性进行综合评价.试验结果表明:橡胶颗粒沥青混合料试件的破坏微应变均超过2 300,满足冬寒区的技术指标;无论是否掺加橡胶颗粒,随着温度的下降,沥青混合料破坏时的最大弯拉强度增大,弯拉应变降低,劲度模量增大;弯曲应变能密度在胶粒掺量为1%左右时具有较大的弯曲应变能密度值,此时橡胶颗粒沥青混合料具有较好的低温抗裂性.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Quantitative trait loci （QTLs） for quality traits related to protein and starch in wheat

Quality traits in wheat （Triticum aestirum L.） were studied by quantitative trait locus （QTL） analysis in a recombinant inbred line （RIL） population, a set of 131 lines derived from Chuan 35050 × Shannong 483 cross （ChSh）. Grains from RILs were assayed for 21 quality traits related to protein and starch. A total of 35 putative QTLs for 19 traits with a single QTL explaining 7.99-40.52% of phenotypic variations were detected on 10 chromosomes, 1D, 2A, 2D, 3B, 3D, 5A, 6A, 6B, 6D, and 7B. The additive effects of 30 QTLs were positive, contributed by Chuan 35050, the remaining 5 QTLs were negative with the additive effect contributed by Shannong 483. For protein traits, 15 QTLs were obtained and most of them were located on chromosomes 1 D, 3B and 6D, while 20 QTLs for starch traits were detected and most of them were located on chromosomes 3D, 6B and 7B. Only 7 QTLs for protein and starch traits were co-located in three regions on chromosomes 1D, 2A and 2D. These protein and starch trait QTLs showed a distinct distribution pattern in certain regions and chromosomes. Twenty-two QTLs were clustered in 6 regions of 5 chromosomes. Two QTL clusters for protein traits were located on chromosomes 1D and 3B, respectively, three clusters for starch traits on chromosomes 3D, 6B and 7B, and one cluster including protein and starch traits on chromosome 1D.