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1.
家禽的性别一般可以通过翻肛法或其他常规方法进行。翻肛方法对个体的应激较大,而且鹅、鸽等家禽进行翻肛鉴定错误率较高。本研究针对禽类性染色体上的染色质解螺旋蛋白DNA结合蛋白1(chromodomain helicase DNA binding protein 1,CHD1)基因,设计了一对引物g CHD,对鸡(Gallus gallus)、鸭(Anas platyrhynchos)、鹅(Anser anser)和鸽(Columba livia)的性染色体CHD1基因进行PCR扩增。扩增结果显示,雄性禽类只有一条带,雌性禽类有两条带,其中较长的带与雄性的条带位置相同。对扩增产物进行克隆测序,序列分析结果表明,较长的条带序列位于Z染色体,命名为CHD1-Z条带;较短的条带序列位于W染色体,命名为CHD1-W带。在鉴定的4个禽类物种中,CHD1-Z条带与CHD1-W条带大小相差均为150 bp左右,其缺失的序列位于内含子中。结果表明,设计的g CHD引物可以作为禽类的通用引物,通过扩增产物凝胶电泳后的条带数目可快速、准确地鉴定禽类的性别。该方法可以提前鉴定出禽类的性别,提高养殖效率;而且为特定性别的胚胎研究提供了可靠的性别鉴定手段。 相似文献
2.
半滑舌鳎性别特异微卫星标记的SCAR转化及其应用 总被引：3，自引：0，他引：3
半滑舌鳎(Cynoglossus semiliaevis)性别控制与全雌育种相关研究需要一种快速、准确的性别特异分子标记。本研究通过对半滑舌鳎共显性性别特异微卫星标记筛选,得到Z、W染色体同源片段相差35bp的位点,对其进行克隆和测序,针对所得测序结果,重新设计引物进行序列特异扩增区间(sequence characterized amplified region,SCAR)转化,增大35 bp差异片段在扩增片段中的所占比例;并摸索得到4%琼脂糖凝胶,150 V,25 min为最适电泳条件,所得SCAR标记可在雌性个体中扩增出169和134 bp两种DNA条带,雄性个体中扩增出169 bp DNA条带,超雌个体中扩增出134 bp DNA条带;将其应用于生产实践中,对半滑舌鳎生理雄性亲鱼576尾进行快速检测,在保证存活的前提下,剔除了亲鱼中的100尾伪雄鱼,有助于提高后代的雌性比例。本研究所得半滑舌鳎性别特异SCAR标记,可用于实验室对半滑舌鳎雌性、雄性和超雌个体遗传性别的准确测定和养殖场简易环境下养殖亲鱼雄鱼中伪雄个体的快速检测。 相似文献
3.
利用羽毛对鸽子进行分子性别鉴定 总被引：2，自引：0，他引：2
鸽子是一种单态鸟,并且是典型的"一夫一妻制",给配对和人工繁殖带来了很大的困难.以雏鸽(Columbalivia Gmelin)的羽毛为材料,采用PCR方法扩增染色体螺旋蛋白基因.结果显示,雏鸽羽毛可以提取出高质量的DNA,雄鸽仅有CHD-Z(450 bp)1条带,而雌鸽有CHD-W(380 bp)和CHD-Z(450 bp)2条带.这种性别鉴定技术准确,并且对动物无伤害性,在生产管理上有广阔的应用前景. 相似文献
4.
半滑舌鳎(Cynoglossus semiliaevis)遗传性别快速准确鉴定的方法是性别控制与高雌苗种培育的基础,其性别决定机制为ZW型,遗传雌鱼为异型染色体ZW,遗传雄鱼为同型染色体ZZ.本研究在半滑舌鳎全基因组测序结果的基础上,通过分析Z染色体和W染色体差异序列,设计一对跨Z/W染色体同源差异DNA片段的引物CS-SEX-F和CS-SEX-R,取半滑舌鳎部分鳍条通过碱煮沸方法获得的DNA为模板,采用PCR扩增,琼脂糖凝胶电泳方法鉴定半滑舌鳎遗传性别.通过该方法在遗传雄鱼(ZZ)个体中可以扩增出366 bp的DNA条带,遗传雌鱼(ZW)个体中可以扩增出366和253 bp两种DNA条带.本研究建立了一种新的成本低廉、操作快速、结果准确可靠的鉴定半滑舌鳎遗传性别的方法,该方法与已有方法相比,操作简便省时,为半滑舌鳎遗传学研究与性别控制育种相关研究提供了可靠分子标记,在半滑舌鳎遗传性别鉴定和养殖场所内伪雄鱼快速检测方面具有广阔的应用前景. 相似文献
5.
睾丸特异蛋白基因对奶牛早期胚胎性别的鉴定 总被引：1，自引：0，他引：1
为方便性别鉴定方法在现场应用，利用睾丸特异蛋白基因（TSPY）建立奶牛早期胚胎的非电泳性别鉴定方法。本实验首先设计并合成TSPY雄性特异引物和雌、雄共有引物并利用已知性别的奶牛血液DNA检测了利用TSPY基因鉴定胚胎性别的可能性。结果显示：雄性特异引物和共有引物对在10pg～60pg范围内性别符合率均为100%；采用非电泳的方法检测TSPY基因鉴定了49枚胚胎的性别，同时，用LAMP法对其中的6枚胚胎的性别鉴定结果进行验证，结果二者完全一致；对其中鉴定为雌性的21枚进行移植，结果9头出生的犊牛均为母牛。结果表明，TSPY基因是一个很好的雄性特异标记，非电泳检测TSPY基因对奶牛早期胚胎性别的鉴定结果准确可靠。 相似文献
6.
为方便性别鉴定方法在现场应用,利用睾丸特异蛋白基因(TSPY)建立了奶牛早期胚胎的非电泳性别鉴定方法.设计并合成了TSPY雄性特异和雌、雄共有基因引物,并利用已知性别的奶牛血液DNA为模板,初步建立了性别鉴定的PCR反应体系和非电泳的性别鉴定方法.灵敏度试验结果显示,雄性特异引物和共有基因引物对在模板10～60 Pg时,其性别鉴定的准确率为100%,提示TSPY基因具备鉴定胚胎性别的可能;同时采用非电泳法和环介导的等温扩增(LAMP)法鉴定6枚胚胎的性别,两者结果完全一致;用非电泳法检测TSPY基因鉴定了43枚胚胎的性别,对其中鉴定为雌性的21枚胚胎分别移植给自然发情后6～8天的受体,结果9头出生的犊牛均为母牛.结果表明.TSPY基因是一个很好的雄性特异标记,非电泳检测TSPY基因对奶牛早期胚胎性别的鉴定结果准确可靠. 相似文献
7.
应用牙釉蛋白(AML)基因鉴别牛早期胚胎性别 总被引：2，自引：1，他引：2
牛牙釉蛋白（AML）基因定位于牛X染色体和Y染色体上的同源区域.实验根据牛X、Y染色体AML基因第五内含子上序列同源性只有45.1%以及X、Y染色体该区段之间存在多处碱基缺失的特点,合成了1对性别特异性引物.该引物经扩增后母牛只得到1条467 bp的来自X染色体的扩增条带,而公牛能得到1条源自X染色体的467bp片段的扩增条带和1条源自Y染色体的341 bp的扩增条带.经基因组DNA验证,该引物的公母鉴定准确率为100%,同时该对引物具有高度的牛特异性.定量分析AML基因引物的反应灵敏度,发现1次PCR（30个循环）能扩增出0.5ng的基因组DNA,2次PCR（共50个循环）能扩增出20pg的基因组DNA.将AML基因引物与SRY基因引物的反应灵敏度相比较,结果表明前者的灵敏度比后者的高8～10倍.用AML基因反应体系鉴定了26枚优质胚胎,其中24枚有扩增结果,13枚鉴定为公牛,11枚鉴定为母牛. 相似文献
8.
根据牛、羊Y－染色体性别决定区域（SRY）的同源性设计了一对PCR引物，对8个绵羊胎儿成纤维细胞系SFF1－8进行了性别鉴定。阳性对照和细胞系SFF1、2、6、7扩增得到130bp片段，阴性对照、空白对照及细胞系SFF3、4、5、8没有扩增带。通过序列测定和同源性分析，证明PCR产物为SRY基因片段，说明有130bp扩增带的细胞系为雄性；无扩增带的为雄性。结果表明，该法具有简单、快速、准确的特点，可应用于转基因克隆动物研究中对体细胞系的早期性别鉴定。 相似文献
9.
加强对进口饲料中牛羊源成分的检测是防止疯牛病和痒病传播的一个重要措施。根据已发表的牛和羊特异性基因及引物序列，分别设计了1条牛和羊特异性semi-nested PCR引物，并采用semi-nested PCR技术对饲料中的牛和羊成分进行了扩增检测。结果表明，semi-nested PCR能够扩增得到247 bp的牛特异性基因条带和214 bp的羊特异性基因条带，其对饲料中牛或羊源性成分的检测灵敏度可达到0.00001%～0.0001%，比普通PCR检测灵敏度要高出103倍；对牛或羊成分DNA的检测灵敏度可以达到10-6～10-5 ng，比普通PCR检测灵敏度要高出105倍以上。该技术具有快速、灵敏和结果稳定的特点，是检测饲料中痕量牛羊源成分的一种有效方法。 相似文献
10.
为了能在分子水平上有效鉴定具有粘果山羊草(Aegilops kotschyi)、偏凸山羊草(A.ventricosa)、普通小麦变种斯卑尔脱(Triticum spelta)细胞质雄性不育系及其保持系90-110和8222,提高其在杂交小麦(Triticum aestivum L.)研究与应用中的定向遗传改良,本研究对其线粒体DNA进行了扩增片段长度多态性(amplified fragment length polymorphism,AFLP)标记和序列特征性片段扩增区域(sequence characterized amplified region,SCAR)标记。通过AFLP标记方法,应用64对引物组合EcoRⅠ-NNN/MseⅠ-NNN对小麦同核异质雄性不育系和保持系进行扩增,共扩增出682条带,其中113条为多态性条带。引物E-AGG/M-CTA组合在粘果山羊草细胞质雄性不育系中扩增出一条大小约300 bp的特异性条带,对该特异条带进行回收、测序,利用Primer Premier 5.0软件重新设计SCAR引物,并对这3种类型同核异质小麦细胞质雄性不育系和保持系进行扩增,其中引物YW1在3种细胞质雄性不育系和保持系中都扩增出条带,而引物YW2仅在粘果山羊草细胞质雄性不育系扩增出一条198 bp的特异性片段,结果表明,已成功地将AFLP标记转化为操作简便、表现稳定的SCAR标记。此片段与小麦线粒体基因组有很高的同源性(同源性为99%),为烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶基因(GenBank登录号:EU534409.1)上的序列,该酶是线粒体中氧化磷酸化的入口酶,与小麦细胞质雄性不育密切相关。本研究可以用于粘类小麦细胞质雄性不育系分子标记辅助育种,也为小麦细胞质种性鉴定提供了技术支撑和理论依据。 相似文献
11.
The number of oocytes in the ovaries of Archispirostreplus tumuliporus judaicus, and the clutch size were determined in two populations of millipedes: one from a mesic habitat and one from a xeric habitat. The presence of males was found to be essential for egg-laying by the females. Daylength did not affect the onset of egg-laying or the clutch size, whereas soil moisture conditions affected egg-laying. Dry soil did not stimulate egg-laying in the xeric-inhabiting females, and caused a decline in clutch size in the mesic ones. Females from the xeric habitat when kept in the laboratory “preferred” laying eggs in the moist soil when given a choice, regardless of the depth at which the soil was moist. 相似文献
12.
肉鸽规模化养殖是一种具有较高经济效益的新兴养殖产业,人工饲喂工作量大、饲喂精细化水平低、饲料浪费率高。为解决肉鸽工厂化规模养殖的自动饲喂难题,设计了一种肉鸽自动饲喂装置。该装置主要由饲喂食槽、控制箱、行程开关、三相异步电机、行走轮、动力传动系统及机架等组成;控制系统使用变频器控制2台三相异步电机的转速,以行程开关为位置检测元件,用循环时间继电器设定饲喂过程中的行走、停留时间,实现自动饲喂装置工作过程中的行走、停留、反向等行程控制,以保证饲喂的精细程度。试验结果表明,该装置行程精度控制在98%以上,饲料浪费率控制在1%以下,具有良好的稳定性,适合于肉鸽工厂化规模养殖。肉鸽自动饲喂装置适用于肉鸽大规模、工厂化养殖。 相似文献
13.
R. Brand Phillips Brian D. Cooke Victor Carrión Howard L. Snell 《Biological conservation》2012,147(1):264-269
Alien species can negatively affect global biodiversity, especially on islands. Significant advances in methods for eradicating mammals from islands have been achieved. In contrast, development of methods for eradicating birds from islands has lagged and few islands have had alien birds successfully eradicated. We report on a 7 year campaign to remove rock pigeons from the Galapagos Islands. To date this is the largest successful eradication of an alien bird from an island system and the only eradication of rock pigeons from an island. Multiple methods were tested and used, including alpha-chloralose, however, shooting with a high-powered air rifle was the most efficient removal technique. Incorporating the support of the community and local agencies into the campaign was critical to the success of the project. 相似文献
14.
Proteomic analysis of wheat flour allergens 总被引：2，自引：0，他引：2
Akagawa M Handoyo T Ishii T Kumazawa S Morita N Suyama K 《Journal of agricultural and food chemistry》2007,55(17):6863-6870
Wheat can cause severe IgE-mediated systematic reactions, but knowledge on relevant wheat allergens at the molecular level is scanty. The aim of the present study was to achieve a more detailed and comprehensive characterization of the wheat allergens involved in food allergy to wheat using proteomic strategies, referred to as "allergenomics". Whole flour proteins were separated by two-dimensional gel electrophoresis with isoelectric focusing and lithium dodecyl sulfate-polyacrylamide gel electrophoresis. Then, IgE-binding proteins were detected by immunoblotting with sera of patients with a food allergy to wheat. After tryptic digestion, the peptides of IgE-binding proteins were analyzed by matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry. In this study, we identified four previously reported wheat allergens or their sequentially homologous proteins [serpin, alpha-amylase inhibitor, gamma-gliadin, and low molecular weight (LMW) glutenin] by a database search. As a result of the high resolution of two-dimensional gel electrophoresis, nine subunits of LMW glutenins were identified as the most predominant IgE-binding antigens. The two-dimensional allergen map can be beneficial in many ways. It could be used, for example, for precise diagnosis of wheat-allergic patients and assessment of wheat allergens in food. Additionally, we compared allergenomics to conventional biochemical methods and evaluated the usefulness of a proteomic strategy for identifying putative allergens to wheat allergy. 相似文献
15.
Identification of collembolan species is generally based on specific morphological characters, such as chaetotaxy and pigmentation pattern. However, some specimens do not match to described characters because these refer to adult specimens, often of one specific sex, or the characters are highly variable in adults (e.g. pigmentation, setae or furcal teeth). Isozymes have frequently assisted species discrimination, and also these may vary with developmental stage or environmental conditions. For identification of single species of the Isotoma viridis group, we present both direct sequencing of the cytochrome oxidase subunit II (COII) gene and a simple DNA-based molecular method.
Five PCR primers amplifying the COII region (717 bp) of the mitochondrial DNA were used. The sequences clearly separated the species I. viridis, I. riparia and I. anglicana, irrespective of colour varieties within the first species. DNA amplification products of different species can also be distinguished by digestion with restriction endonucleases, followed by gel electrophoresis for separation of fragments. This restriction fragment length polymorphism (RFLP), obtained after digestion with the endonucleases TaqI, VspI, MvaI and Bsp143I, revealed specific fragments that separated the three species from each other. Since restriction enzymes are sensitive to single base mutations, we suggest to use a combination of enzymes with at least two species-specific restriction sites when using the RFLP technique. For the I. viridis complex, VspI and Bsp143I appear to be an appropriate combination. 相似文献
16.
为了分析不同饲养模式和阶段对蛋鸡发声的影响,并为构建基于蛋鸡声音信息的健康养殖评价系统提供参考,该研究对蛋鸡的发声进行了监测,通过声音预处理、特征提取、数据挖掘和统计分析等方法,研究笼养和栖架饲养模式下、育成期和产蛋期蛋鸡的声学特征。结果表明,典型蛋鸡声音可分为产蛋叫声、鸣唱声、鸣叫声和争斗尖叫声等四类。产蛋期蛋鸡发声的峰值频率和声音能量水平均低于育成期蛋鸡。同时发现蛋鸡发声的峰值频率与蛋鸡周龄大小呈现负相关关系,由14周龄的(2 192±320)Hz降至41周龄的(1 550±345)Hz。比较笼养和栖架养殖模式下蛋鸡的声音特征发现:栖架养殖模式下蛋鸡发出的声音信号次数、持续时间和声音能量均高于笼养模式,栖架养殖下的蛋鸡发声数量是笼养模式下蛋鸡的3倍以上,栖架系统内蛋鸡白天的声音能量比笼养蛋鸡高接近1倍,这些结果表明在以福利化为目标的栖架养殖模式中蛋鸡表达更多的自然行为,蛋鸡发声的丰富程度可用于后续开发评价蛋鸡的福利状况的方法。 相似文献
17.
本实验以16个石榴品种为实验材料,筛选出10个重复性及多态性均较好的引物进行RAPD分析。分别采用琼脂糖凝胶以及聚丙烯酰胺凝胶(PAGE)电泳检测方法对PCR扩增结果进行检测并对其结果进行比较,结果显示,两种电泳方式均能得到较为清晰的扩增条带,且两种电泳方式获得的条带总数及多态性条带数均有所不同,琼脂糖凝胶电泳方法共检测出76条带,其中有43条为多态性谱带,多态性比率为56.4%;而在PAGE电泳方法共检测出123条谱带,多态性谱带数为87条,多态性比率为70.95%。PAGE电泳方法检测出的条带数约为琼脂糖凝胶电泳方法检测出条带数的1.5倍。基于两种电泳方法所得RAPD标记的多态性位点,利用NYSYS软件计算遗传相似系数,并构建遗传关系聚类图,分析结果显示,石榴遗传多样性丰富,两种电泳方法所得聚类结果大致相同,可以利用RAPD分子标记及两种电泳检测方法对不同数量的石榴进行分子水平的品种鉴定和遗传多样性的分析。同时通过对来自几个引物随机挑选的17个片段进行克隆,测序结果显示17个片段都是对应引物的RAPD扩增产物,其中有3条是编码蛋白的基因片段,表明了RAPD不仅扩增基因组上的非编码蛋白序列,同时也可以扩增编... 相似文献
18.
利用双向电泳分离棉铃虫中肠Cry1Ac受体蛋白,对于进一步明确棉铃虫受体的种类、发现新受体、寻找可能与抗性有关的受体非常重要。根据第一向等电聚焦的方式不同,可将双向电泳分为两种系统：ISO-DALT(isoelectric focus-dalton weight)和IPG-DALT（immobilized pH gradient-dalton weight）。确定最适合的双向电泳方法,可以为进一步研究受体蛋白奠定基础。本研究比较了ISO-DALT和IPG-DALT两种双向电泳系统结合western blotting技术分离棉铃虫中肠Cry1Ac受体蛋白的效果。结果表明,这两种系统都可以用于棉铃虫Cry1Ac受体的分离,ISO-DALT对受体分离效果比IPG-DALT更好,ISO-DALT比较经济但对后续的质谱工作不利,IPG-DALT较昂贵但有利于进行后续试验。 相似文献
19.
Ilse Storch 《Biological conservation》1994,70(3):237-243
The capercaillie Tetrao urogallus has become endangered in central Europe, most probably as a result of habitat changes. Breeding ecology was studied by following radio-tagged hens through egg-laying, incubation, and chick-rearing between 1988 and 1992. Loss of chicks rather than nests was the main determinant of reproductive success. All adult hens incubated, two-thirds of the nests hatched, and one-fifth of the chicks survived until autumn. Both nests and broods were mostly found in habitats with rich ground vegetation. Nest cover was probably a major factor for hatching success. Brood home ranges averaged 148 ha during the period from hatching to late summer. Broods preferred old forest stands with rich ground vegetation and high invertebrate abundance, which occur under moderate canopy cover. Bilberry Vaccinium myrtillus was an essential feature for brood habitat at forest stand, home range, and landscape scale, and should therefore be a major goal for capercaillie habitat management in central Europe. 相似文献
20.
Muroya S Ohnishi-Kameyama M Oe M Nakajima I Shibata M Chikuni K 《Journal of agricultural and food chemistry》2007,55(10):3998-4004
To investigate changes in myosin light chains (MyLCs) during postmortem aging of the bovine longissimus muscle, we performed two-dimensional gel electrophoresis followed by identification with matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The results of fluorescent differential gel electrophoresis showed that two spots of the myosin regulatory light chain (MyLC2) at pI values of 4.6 and 4.7 shifted toward those at pI values of 4.5 and 4.6, respectively, by 24 h postmortem when rigor mortis was completed. Meanwhile, the MyLC1 and MyLC3 spots did not change during the 14 days postmortem. Phosphoprotein-specific staining of the gels demonstrated that the MyLC2 proteins at pI values of 4.5 and 4.6 were phosphorylated. Furthermore, possible N-terminal region peptides containing one and two phosphoserine residues were detected in each mass spectrum of the MyLC2 spots at pI values of 4.5 and 4.6, respectively. These results demonstrated that MyLC2 became doubly phosphorylated during rigor formation of the bovine longissimus, suggesting involvement of the MyLC2 phosphorylation in the progress of beef rigor mortis. Keywords: Bovine; myosin regulatory light chain (RLC, MyLC2); phosphorylation; rigor mortis; skeletal muscle. 相似文献