耐氟喹诺酮类病原菌gyrA基因突变的寡核苷酸芯片检测

根据大肠杆菌、沙门菌、无乳链球菌和鸡毒霉形体的gyrA基因序列，设计了通用引物和ll条寡核苷酸探针；利用点样仪将探针点在基片上，制成寡核苷酸芯片；采用PCR荧光标记靶基因，与芯片杂交，用荧光扫描仪检测信号；同时以PCR一测序法进行gyrA基因突变的检测。结果，PCR反应体系能特异性地扩增出靶基因；寡核苷酸芯片能同时检测不同病原菌GyrA第83、87位发生的突变，芯片检测结果与测序结果较为一致。结果表明，使用寡核苷酸芯片技术检测病原菌耐氟喹诺酮类基因突变是可行的；研究结果为基因芯片技术应用于兽医临床耐药性检测提供了基础。     

一种提取荒漠拟步甲昆虫基因组DNA的新方法

昆虫基因组DNA的提取是研究昆虫功能基因的关键环节。一些荒漠昆虫个体较小,不易除去的外壳会造成多糖污染,用普通动物组织DNA提取方法效果不佳。通过紫外分光光度测定、凝胶电泳检测、PCR反应及限制性酶切分析表明,CTAB-NaCl法是一种适用于拟步甲科小个体昆虫提取基因组DNA的好方法,蛋白质、多糖及RNA影响很低,完全能够满足后续DNA研究的需求。与传统方法相比简便快速,且成本低,尤其适用于拟步甲科昆虫DNA的分离。     

跳枝碧桃花色性状的全基因组关联分析

跳枝碧桃(Prunus persica f. versicolor)在同一植株上可产生二色花和嵌合花,具有很高的观赏价值。以南京3个观赏桃集中分布区的57株跳枝碧桃为材料,利用简单重复序列(SSR)标记研究其遗传背景。通过PCR扩增,筛选出6对多态性引物,共计扩增出17个多态性条带,平均有效等位基因2.83个。SSR标记基因分型结果表明,供试材料可分为3种基因型,各采样地均具有2～3种基因型,同一植株的不同分枝基因型完全一致。基于基因分型结果,选取3种基因型跳枝碧桃各1株进行基因组重测序,结合公开发表的数据,进行位点变异分析,并构建系统发育树。结果显示3种基因型的跳枝碧桃聚在一个分支,其中B20、T3基因型与‘洒红桃’聚在一起,W5基因型相对独立。3种基因型的跳枝碧桃在起源上比较接近,但不同基因型之间存在一定的遗传差异。通过整合单核苷酸多态性(SNP)信息和花色表型开展全基因组关联分析(GWAS),鉴定出5个SNP与花色跳枝性状显著关联(P值<10^-100),均位于7号染色体。在显著SNP位点的邻近基因组区段发掘3个DICER-LIKE2(DCL2)基因,揭...     

基因组选择在家畜改良上的研究进展

基因组选择（GS）是全基因组范围内的分子标记辅助选择,目前被证明是利用DNA标记信息改善复杂性状最有效的方法。本文简要概述了基因辅助选择以及标记辅助选择;重点介绍了GS,包括GS的实施策略与育种值估计方法,GS的准确性获得以及对GS方法的比较,总结了当前家畜上利用GS加速遗传改良的应用进展;并对家畜在GS上的应用前景进行展望。     

Genome‐wide association study for feedlot average daily gain in Nellore cattle (Bos indicus)

The genome‐wide association study (GWAS) results are presented for average daily gain (ADG) in Nellore cattle. Phenotype of 720 male Bos indicus animals with information of ADG in feedlots and 354 147 single‐nucleotide polymorphisms (SNPs) obtained from a database added by information from Illumina Bovine HD (777 962 SNPs) and Illumina BovineSNP50 (54 609) by imputation were used. After quality control and imputation, 290 620 SNPs remained in the association analysis, using R package Genome‐wide Rapid Association using Mixed Model and Regression method GRAMMAR‐Gamma. A genomic region with six significant SNPs, at Bonferroni‐corrected significance, was found on chromosome 3. The most significant SNP (rs42518459, BTA3: 85849977, p = 9.49 × 10^?8) explained 5.62% of the phenotypic variance and had the allele substitution effect of ?0.269 kg/day. Important genes such as PDE4B, LEPR, CYP2J2 and FGGY are located near this region, which is overlapped by 12 quantitative trait locus (QTLs) described for several production traits. Other regions with markers with suggestive effects were identified in BTA6 and BTA10. This study showed regions with major effects on ADG in Bos indicus in feedlots. This information may be useful to increase the efficiency of selecting this trait and to understand the physiological processes involved in its regulation.     

Accurate genomic predictions for BCWD resistance in rainbow trout are achieved using low‐density SNP panels: Evidence that long‐range LD is a major contributing factor

Previously accurate genomic predictions for Bacterial cold water disease (BCWD) resistance in rainbow trout were obtained using a medium‐density single nucleotide polymorphism (SNP) array. Here, the impact of lower‐density SNP panels on the accuracy of genomic predictions was investigated in a commercial rainbow trout breeding population. Using progeny performance data, the accuracy of genomic breeding values (GEBV) using 35K, 10K, 3K, 1K, 500, 300 and 200 SNP panels as well as a panel with 70 quantitative trait loci (QTL)‐flanking SNP was compared. The GEBVs were estimated using the Bayesian method BayesB, single‐step GBLUP (ssGBLUP) and weighted ssGBLUP (wssGBLUP). The accuracy of GEBVs remained high despite the sharp reductions in SNP density, and even with 500 SNP accuracy was higher than the pedigree‐based prediction (0.50–0.56 versus 0.36). Furthermore, the prediction accuracy with the 70 QTL‐flanking SNP (0.65–0.72) was similar to the panel with 35K SNP (0.65–0.71). Genomewide linkage disequilibrium (LD) analysis revealed strong LD (r² ≥ 0.25) spanning on average over 1 Mb across the rainbow trout genome. This long‐range LD likely contributed to the accurate genomic predictions with the low‐density SNP panels. Population structure analysis supported the hypothesis that long‐range LD in this population may be caused by admixture. Results suggest that lower‐cost, low‐density SNP panels can be used for implementing genomic selection for BCWD resistance in rainbow trout breeding programs.     

Association of three RNA molecules with potato mop-top virus

Summary Potato mop-top virus particles, purified from systemically infectedNicotiana benthamiana plants and then disrupted by heating with sodium dodecyl sulphate and 2-mercaptoethanol, contained only a single polypeptide of M_r 19 100 detectable by polyacrylamide gel electrophoresis. Single-stranded RNA preparations from virus particles, when subjected to electrophoresis in an agarose gel containing methylmercuric hydroxide as a denaturant, were shown to contain approximately equal proportions of three RNAs of 6.5, 3.2 and 2.5 kb. Double-stranded RNA preparations extracted from systemically infectedN. benthamiana leaves or from locally infectedN. debneyi leaves, was shown by polyacrylamide gel electrophoresis to contain a major species of 3.2 kbp and two minor species of 6.5 and 2.4 kbp.     

Alternatives to Blood as a Source of DNA for Large-scale Scanning Studies of Canine Genome Linkages

Participation and compliance are critical to the success of any large-scale study of canine disease using DNA markers. Most canine genetic studies rely upon DNA extracted from peripheral blood samples. We assessed the utility of buccal swab epithelial cells and toe nails as a source of DNA for use in genomic screening studies. Using eight multiplexed canine microsatellite markers, amplified DNA obtained from peripheral blood, and from freshly extracted buccal epithelial cells, and buccal swab DNA extracted and stored at –20°C for 27 months or extracted from toe nails were compared for three dogs. The accuracy of the genotyping at each locus was identical for each preparation. Buccal swab DNA samples were readily and uniformly amplified and could be stored for years without loss of integrity. Each buccal swab provided sufficient DNA for more than 200 individual PCR reactions. Toe nails provided ample DNA for thousands of PCR reactions and had the added advantage of ease of storage of the original tissues. These studies demonstrate the potential utility of DNA derived from buccal swabs or nails in large-scale genomic scanning and marker linkage studies.     

中、西蜂纯种及其不同品系蜂的基因组DNA多态性分析

使用P8(5‘-CCCACCCTTG-3‘)和W2（5‘-CGGCCCCGGT-3‘)随机引物，通过PRAPD（Random Amplified Polymorphic DNA)-PCR(polymerase chain reaction)遗传学分析，分别对中、西纯种蜂的不同品系的基因组DNA进行多态性分析。结果：中、西纯种蜂的不同品系的DNA多态4性图谱均具有明显的差异，从而为中、西纯种蜂的品系之间的鉴别提供了科学依据。