应用实时荧光定量PCR测定牛体细胞端粒长度

本研究为了验证应用实时荧光定量聚合酶链反应（quantitative PCR,Q-PCR）测定牛基因组端粒长度的可行性,选取18个不同来源牛耳成纤维细胞抽提基因组DNA为样本,对Q-PCR和经典的Southern印迹法进行了相关性分析。结果显示,Q-PCR测定端粒长度相对T/S为1.16±0.24,Southern印迹法测量端粒平均TRF值为16.99 kb±0.85 kb,两种方法获得的结果相关性分析R2=0.5612(P＜0.01),因此实时荧光定量PCR是一种测定牛基因组端粒长度的可靠的方法。     

lncRNAs调控表观遗传的研究进展

表观遗传学(epigenetic)是不改变DNA序列而使基因的表达发生可遗传的变化,包括DNA甲基化、组蛋白修饰、基因组印迹、随机染色体失活及长链非编码RNA(lncRNAs)的调节作用等。lncRNAs是一类转录本长度超过200nt的RNA分子,它们并不编码蛋白,而是以RNA的形式在多种层面上调控基因的表达水平。lncRNAs的功能往往受到表观遗传作用的影响,同时lncRNAs通过染色质修饰、基因组印迹、剂量补偿效应等过程,在基因表达中发挥表观遗传学作用。论文对lncRNAs表观遗传调控的研究进展情况进行了综述,以期为深入研究lncRNAs调控性状的机理提供思路。     

动物印迹基因与胚胎发育关系的研究进展

基因印迹是不遵循孟德尔遗传规律的一种依靠单亲传递某些遗传性状的现象,即某些等位基因呈亲缘依赖性表达,另一等位基因不表达或表达极弱.配子和早期胚胎形成期是甲基化印迹擦除,建立和维持的关键窗口期,而此期恰为体外生产胚胎时卵母细胞和胚胎体外培养阶段,甲基化易受体外受精(IVF)、精子显微注射(ICSI)干预,进而干扰基因组印...     

应用泛特津试剂盒提取染色体DNA的方法

采用一种简单、快速、安全、无污染的方法，从黑曲霉细胞中提取基因组DNA，以满足各种目的的PCR、Southern印迹的分析要求。该方法可在短时间内制备大量样品，浓度高、质量好，操作简单，实用，推广性强。     

DNA甲基化与基因沉默

DNA甲基化是生物体基因组的主要表观遗传修饰方式,是调节基因组功能的重要手段,哺乳动物通过基因组甲基化来改变DNA与蛋白质的作用,提供非编码序列和与发育相关的基因沉默的可遗传机制.     

抗菌肽Thanatin转基因小鼠建立的初步研究

人工合成抗菌肽Thanatin基因的3条片段,利用重叠延伸PCR扩增技术得到完整的Thanatin基因,通过分子克隆方法构建出pIRES2-EGFP-Thanatin重组表达载体。然后与脂质体混合,采用睾丸打点注射将新构建的抗菌肽基因注入小鼠睾丸内,共注射公鼠5只。6周后与雌鼠交配,对新生仔鼠进行断尾,提取尾尖基因组DNA,利用PCR和Southern印迹方法对其进行检测。结果显示,得到的52只新生仔鼠中PCR检测阳性率为38.46%,Southern印迹检测阳性率为30.77%;在活体荧光成像系统下转基因小鼠呈现绿色荧光表达。通过睾丸注射法使Thanatin基因在小鼠的基因组中得到整合,为进一步研究抗菌肽的作用机理、培育抗病动物品种以及通过建立转基因动物生物反应器进行抗菌肽的大量生产奠定了基础。     

表观遗传学与表型变异

<正>鸡的基因组是农业领域首批被测序和注释(尽管不完全)的动物基因组之一,其特点是基因组相对较小、存在表观遗传属性迥异的微型和大型染色体以及哺乳动物不同的性别决定方式。虽然对鸡来说,表观遗传机制(如DNA甲基化、组蛋白修饰和染色质调控)看似保守,但某些机制与哺乳动物相比可能有所不同。     

农杆菌介导白三叶草高效遗传转化和转基因植株再生

利用子叶下胚轴为外植体,通过农杆菌Ti质粒介导途径,建立了白三叶草高效遗传转化及转基因植株高频率再生体系。PCR检测及Southern印迹鉴定结果表明,外源T-DNA片段已整合到转基因植株的基因组中,且多以1~2个少数拷贝存在。Northern印迹结果和对报告基因GusA编码蛋白的活性检测结果表明,目的基因在部分转基因植株中高水平表达。适宜条件下外植体遗传转化后的植株再生比例达58.23%~62.12%。与对照相比,转基因植株在外部形态上没有发生改变。     

AnnualReviewofEntomology：家蚕基因组序列解析推动了家蚕的前沿研究

自从2003年家蚕全基因组框架图测序计划完成以来，基于基因组序列的家蚕比较基因组学与功能基因组学研究得到了极大的推动。近10年来，中国西南大学家蚕基因组生物学国家重点实验室领衔先后完成了家蚕全基因组精细图、全基因组遗传变异图及转录组图谱构建，并重点开展了家蚕丝蛋白合成及免疫抗性等性状形成的分子基础解析；中国科学院上海植物生理生态研究所及华南师范大学长期致力于家蚕变态发育的分子调节机制研究。     

H5N1亚型禽流感病毒进化的研究进展

H5N1亚型禽流感病毒是重要的人兽共患病病原,其基因组为分节段的负股单链RNA,这种特殊的基因组结构使得流感病毒的进化机制跟其他病毒有所不同。本文对流感病毒的进化机制、H5N1亚型禽流感病毒的进化规律和生物信息学在流感病毒进化中应用等方面的研究进展予以综述。     

Study on the mechanism of maternal imprinting during oocyte growth

In mammals, both parental genomes are essential for normal ontogeny because epigenetic modifications imposed in the parents' gametes lead to parent-of-origin specific gene expression in their offspring. These phenomena are referred to as genomic imprinting. It has been shown that maternal imprinting is established during oocyte growth, lack of maternal imprinting in zygotes leads to early embryonic death, and in vitro system that allows establishment of maternal imprinting is developed. In this review, I describe the history of the discovery of genomic imprinting, the regulatory mechanisms of mammalian development by maternal imprinting, and the molecular mechanisms of genomic imprinting.     

基因组印记的研究进展

综述基因组印记的可能发生机制及人和鼠中常见的印记基因，论述了且印记作用的生物学意义。     

家畜基因印记研究进展

综述了家畜基因印记的生理功能、可能机制及家畜中发现的印记基因,并论述了基因印记的生物学意义和对家畜育种的影响。     

Genomic imprinting variances of beef carcass traits and physiochemical characteristics in Japanese Black cattle

The objective of this study was to estimate variance components related to imprinting for carcass traits and physiochemical characteristics in Japanese Black cattle. The carcass records obtained from 4,220 Japanese Black feedlot cattle included carcass weight (CW), rib eye area (REA), rib thickness, subcutaneous fat thickness, and beef marbling score (BMS), and the physiochemical characteristics were fat, moisture, glycogen per proportion of moisture content, oleic acid, and monounsaturated fatty acids (MUFA). To detect gametic effects, an imprinting model was fitted. High additive heritabilities were estimated for all traits (from 0.516 for glycogen to 0.853 for fat) and were reduced in Mendelian heritability. The range of the differences was from 0.002 (CW) to 0.331 (fat and moisture), and the reductions were due to their imprinting variances. The ratio of the imprinting variance to the total additive genetic variance for REA (0.374), BMS (0.291), fat (0.387), moisture (0.388), and MUFA (0.337) were large (p < 0.05). These imprinting variances were due to the maternal contribution and suggested the existence of maternally expressed genomic imprinting effects on the traits in Japanese Black cattle. Therefore, maternal gametic effects should be considered in breeding programs for Japanese Black cattle.     

Parent‐of‐origin effects on carcass traits in Japanese Black cattle

Variances caused by the differential expression of paternally and maternally imprinted genes controlling carcass traits in Japanese Black cattle were estimated in this study. Data on marbling score (BMS), carcass weight, rib thickness, rib‐eye area (REA) and subcutaneous fat thickness (SFT) were collected from a total of 13,115 feedlot steers and heifers in a commercial population. A sire–maternal grandsire model was used to analyse the data, and then, imprinting parameters were derived by replacing the genetic effect of the dam with the effect of the maternal grandsire in the imprinting model to calculate the genetic parameter estimates. The proportions of the total genetic variance attributable to imprinted genes ranged from 8.7% (SFT) to 35.2% (BMS). The remarkably large imprinting variance of BMS was mainly contributed by maternally expressed inheritance because the maternal contribution of the trait was much larger than that of the paternal trait. The parent‐of‐origin effect originating from maternal gene expression was also observed for REA. The results suggested the existence of genomic imprinting effects on the traits of the Japanese Black cattle. Hence, the parent‐of‐origin effect should be considered for the genetic evaluation of Japanese Black cattle in breeding programmes.     

长链非编码RNA在肌肉发育中的作用研究进展

长链非编码RNA(long noncoding RNA, lncRNA)是一类长度大于200个核苷酸且不具有蛋白编码功能的RNA转录本,研究表明它参与了基因组印记、转录激活与干扰、转录后调控、染色体剂量补偿效应、发育调控等众多生物过程。作者就lncRNA的发现和分类、生物学功能及与肌肉发育相关lncRNA等研究进展进行了综述。     

长链非编码RNA及其在医学、家畜方面的研究进展

长链非编码RNA(long non-coding RNA, LncRNA)是一类转录本长度超过200 nt的RNA分子,没有或少有编码蛋白的能力。LncRNA参与编码基因表达调控的表观遗传机制,能够直接调控靶基因的转录与蛋白的降解等,在基因组印记、转录调控及人类疾病等方面有着广泛功能。作者着重对LncRNA的特点、分类、作用机制及其在医学和家畜方面的研究进展作一综述,最后对LncRNA在家畜方面的研究与应用进行展望。     

早期胚胎DNA甲基化研究进展

DNA甲基化修饰是研究最多的表观遗传修饰之一，在调控基因转录、染色体结构稳定、基因印迹、X染色体失活等方面发挥作用。尽管DNA甲基化是一种稳定的修饰，但其在个体发育进程中是动态变化的。目前，人们对早期胚胎发育中DNA甲基化修饰研究还不全面，随着全基因组DNA甲基化分析技术的进步，其在早期胚胎中的功能也逐渐揭示。作者主要论述了DNA甲基转移酶（DNMTs）的发现及其调控作用和DNA甲基化在早期胚胎中的作用。     

Genomic imprinting effects on dairy- and fitness-related traits in cattle

Data from the official milk recording scheme of the Austrian Simmental population were analysed to estimate variance components due to genomic imprinting. Traits regarded were milk yield, fat and protein content, persistency, days open (first, second and third lactation) and herd life. All dairy traits were preadjusted for BLUP herd‐year effects. After applying some data restrictions, the number of records ranged from 3391 (persistency, third lactation) to 33 993 (days open, first lactation). Two different estimation approaches were used: (i) estimation of the maternal and paternal gametic component by a dam and sire model, respectively, and (ii) estimation of the animal and an additional gametic component (maternal or paternal) by treating gametes as homozygous diploid individuals. All models also accounted for year of first calving and the cytoplasmic effect of maternal lineages. In tendency, the results of both approaches were fairly well in line with each other. For the majority of the investigated traits, no substantial imprinting effects were detected. Significant evidence of genomic imprinting on a multiple type I error ≤ 0.10 was only found for protein content and days open. For protein content in the second and third lactation, the proportions of variance due to the paternal gametic effect exceeded those of the maternal by 0.096 and 0.152, respectively. For days open in the second lactation, however, the proportion of variance of the maternal gamete effect exceeded that of the paternal by 0.036. In tendency, indication of paternal imprinting was found for all fitness‐related traits. For true and functional herd life, significant differences (pairwise type I error ≤ 0.05) of 0.040 and 0.032 were found between the proportions of variance due to the maternal and paternal gamete effect. Significant variance components of cytoplasmic effects (multiple type I error ≤ 0.01) were found for first lactation of milk yield (0.020), for first, second and third lataction of persistency (0.026, 0.035 and 0.033) and of days open (0.029, 0.016 and 0.022) and for true and functional herd life (0.019 and 0.029).