共查询到19条相似文献，搜索用时 343 毫秒
1.
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
2.
为确定合适的亲水胶体种类及其添加量,以带鱼鱼糜为原料,通过测定凝胶强度、质构(硬度、粘聚性、弹性和咀嚼性)、白度、持水性和凝胶溶解度等指标,分析瓜尔胶、魔芋胶和沙蒿胶3种亲水胶体对带鱼鱼糜凝胶品质的影响。结果表明,瓜尔胶、魔芋胶和沙蒿胶均能提高带鱼鱼糜凝胶强度,但相同添加量下魔芋胶的改善效果最好,而瓜尔胶对鱼糜凝胶强度的影响不显著。魔芋胶能显著改进带鱼鱼糜凝胶的质构特性;瓜尔胶添加量为2.0%时鱼糜凝胶硬度、弹性、咀嚼性、粘聚性比对照组低;沙蒿胶对鱼糜凝胶的粘聚性影响显著,当添加量为1.5%或2.0%时,带鱼鱼糜凝胶的咀嚼性、硬度增加,弹性下降。瓜尔胶、魔芋胶和沙蒿胶均能提高带鱼鱼糜持水性,降低鱼糜凝胶白度和溶解度,但对鱼糜凝胶色泽的影响不显著。扫描电镜结果显示,1.5%魔芋胶处理组形成的鱼糜凝胶表面规则有序、网络结构致密均一。综合比较,添加1.5%魔芋胶能有效改善带鱼鱼糜的凝胶品质。本研究结果为提高带鱼鱼糜制品品质及生产研发提供了一定的理论依据。 相似文献
3.
用聚丙烯酰胺凝胶盘状电泳(Polyacrylamide gel disk electrophoresis)法分离血清蛋白,在医学上用来诊断疾病已有不少报导。但在畜牧工作中通过血清蛋白的分离测定来鉴定畜禽品种的报导则不多,尤其是用此法研究广西的一些地区性畜禽的血清蛋白尚未见报导。实验已经证明,动物血清蛋白的合成受遗传基因的控制,不同基因型可生成不同表现型的血清蛋白,不同种类的动物,血清蛋白的种类及含量不同。故本研究采用分辨力高的聚丙烯酰胺凝胶盘状电泳法对广西水牛、广西霞烟鸡与国外引进 相似文献
4.
草菇低温诱导蛋白的分离与纯化 总被引：7，自引：0，他引：7
低温4℃诱导草菇（Volvariella volvacea (Bull.ex Fr)Sing.)菌丝体2h后，对其可溶性蛋白进行分析，发现草菇在低温胁迫下有新的可溶性蛋白质产生。应用硫酸铵分级沉淀和电泳分离制备技术，分离和纯化得到了草菇菌丝体中的一个低温诱导蛋白，用聚丙烯酰胺凝胶电泳和等电聚焦电泳等方法分析该蛋白，结果均呈现为单一条带。SDS－聚丙烯酰胺凝胶电泳结果表明蛋白是由分子量为51kD的一个亚基组成，等电聚焦电泳结果发现该蛋白的等电点为3．73。 相似文献
5.
种子蛋白质电泳在陆地型长绒棉种质鉴定中的应用 总被引：1，自引：0，他引：1
采用垂直平板聚丙烯酰胺凝胶法 ,进行单粒种子醇溶蛋白质电泳分析 ,参试材料有陆地型长绒新种质 9830 1系、4个海岛棉品种和 8个陆地棉品种。 9830 1和中棉所 1 2号、石远 32 1在试验中 ,同一品种 (系 )内的不同种子之间的蛋白谱带基本一致。对分别取样于山东省 7个不同地区试验点的 9830 1种子谱带进行比较 ,没有发现差异。胶板上陆地棉品种均具有 3条与海岛棉相区分的带 ,同样各海岛棉品种也共同具有 3条特征带。 9830 1显出 3条陆地棉特有带 ,证明其谱带主要倾向于陆地棉 ;同时也稳定地出现 1条海岛棉的特有带 ,表现出其属于海陆杂交种的遗传特性。 9830 1分离出一条独有带 (“1”带 ) ,凭借此带和另一条带 (“1 0”带 )可较容易地和其它陆地棉品种相区分 相似文献
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7.
为了进。步分析猪笼草瓶状体内消化液中蛋白质的性质，本研究在不同的温度和pH条件下测定消化液中蛋白酶的活性，并且比较了3种不同沉淀方法对消化液蛋白进行的浓缩，通过SDS-聚丙烯酰胺电泳对消化液蛋白作了初步分离。结果表明，猪笼草消化液中蛋白酶的最适酶活温度为50℃，且稳定性最高；当pH为5时，该蛋白酶活性出现峰值，采用氯仿-正丁醇法（5：1）沉淀浓缩猪笼草消化液蛋白样品效果最佳。聚丙烯酰胺电泳结果表明，消化液至少包括三种蛋白组分，分子量分别为24.3kD、35．1kD和61．4kD，且35．1kD条带具有抗胰蛋白酶消化活性。本研究为综合开发利用猪笼草野生资源提供理论依据。 相似文献
8.
本研究的目的在于筛选合适的RAPD随机引物,应用RAPD技术对药用植物绞股蓝进行遗传多样性分析,并构建DNA指纹图谱。研究结果表明,我们利用生物信息学方法挑选出的20条引物中有19条引物的扩增条带清晰且多态性好;在清晰稳定出现的354条带中,294条具有多态性;其中有3条引物的扩增条带可清楚区分绞股蓝与混淆品种乌蔹莓,可建立其DNA指纹图谱。按UPGMA法进行聚类分析,计算其遗传相似系数,结果显示,8份绞股蓝供试材料聚为两类,聚类结果与其地理区域远近和生长环境一致。本研究中筛选出的19条引物适用于绞股蓝遗传多样性分析,且获得的DNA指纹图谱可用于鉴别绞股蓝。 相似文献
9.
研究旨在鉴定两条在非变性聚丙烯酰胺凝胶电泳中总是伴随杂合子出现的非目的条带.实验切胶回收了目的条带和非目的条带,以此为模板进行第二次PCR扩增,然后将PCR产物按不同组合进行混合并施加不同处理再次进行电泳,另外,人工合成了四条包含该缺失位点的寡核苷酸单链,对条带形成的过程进行了模拟.结果表明以两条非目的条带为模板的第2次PCR产物的电泳结果均有4条带,与杂合子样本4条带的迁移率完全一致,以2条目的条带为模板的第2次PCR产物电泳条带的迁移率分别与其本身保持一致;将以2条目的条带为模板的第2次PCR产物混合并经过变性复性后,其结果出现非目的条带和目的条带,也与杂合子样本4条带的迁移率完全一致,而如果只是简单混合并未经过变性复性处理,其电泳结果只出现目的条带.另外,在利用寡核苷酸单链进行模拟的实验中发现,异源双链迁移率较低,形成了非目的条带,而正常双链迁移率较高,形成目的条带.结果提示,这种与杂合子相伴产生的非目的条带,并不是非特异性扩增的结果,而是由于模板为缺失突变杂合子,在复性过程中,PCR产物中的一部分碱基缺失的正负单链会与不缺失的正负单链发生互补,而在发生碱基缺失的位置,会有突环产生,产生的突环进而导致杂合双链迁移率降低,最终形成了两条迁移率较低的非目的条带. 相似文献
10.
聚丙烯酰胺对蚯蚓的毒性效应 总被引：4，自引：1，他引：3
聚丙烯酰胺作为全球应用最广泛,用量最大的水处理剂,而其排放到环境中可能会对生态环境形成潜在的威胁。该研究在人工土壤条件下,通过急性和亚急性暴露试验研究了聚丙烯酰胺和丙烯酰胺对赤子爱胜蚓存活、生长和繁殖的影响,旨在评价聚丙烯酰胺和丙烯酰胺对蚯蚓的毒性效应。结果表明,聚丙烯酰胺和丙烯酰胺对蚯蚓的半致死剂量分别为大于2000和164.01?mg/kg,聚丙烯酰胺比丙烯酰胺毒性低;在急性和亚急性毒性暴露期内,聚丙烯酰胺对蚯蚓的存活和生长无显著影响;而当丙烯酰胺浓度大于100?mg/kg时即对蚯蚓的存活和生长产生显著的影响（P＜0.05）。聚丙烯酰胺和丙烯酰胺均对蚯蚓的繁殖能力有非常显著的影响（P＜0.05）。因此残留于污泥中的聚丙烯酰胺对环境有一定的潜在风险。 相似文献
11.
Martín-Hernández C Bénet S Obert L 《Journal of agricultural and food chemistry》2008,56(12):4348-4351
Five methods using aqueous/organic solvents for the separation of proteins from oils were compared. The extraction with acetone-hexane followed by amino acid analysis was found to be the most suitable method for isolation and quantification of proteins from oils. The detection limit of the method was 0.18 mg protein/kg oil, and the quantification limit was 0.6 mg protein/kg. The relative repeatability limit for samples containing 1-5 mg protein/kg sample was 27%. The protein recovery ranged between 68 and 133%. Using this method, the protein content of 14 refined and nonrefined oils was determined. In none of the refined oils were proteins detected, whereas the protein content of the unrefined oils ranged between undetectable in extra virgin olive oil to 11 mg/kg in rapeseed oil. With sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with silver staining, many protein bands were visible in the unrefined soy, olive, peanut, and rapeseed oil samples. Proteins bands were not obtained from the refined fish oil. In the other refined oil samples, a few proteins bands could be visualized. Two protein bands with apparent molecular molecular masses of 58 and 64 kDa were always observed in these oils. 相似文献
12.
DGGE法与常规培养法对稻田蓝细菌多态性分析结果比较 总被引：1，自引：0，他引：1
研究运用蓝细菌和硅藻16SrDNA特异引物,将晚季水稻生长后期稻田土壤中提取的总DNA进行PCR扩增后,以DGGE技术对PCR产物进行分析结果表明,14条DGGE带经克隆测序,经NCBI基因库比对得晚季水稻生长后期存在10种蓝细菌,包括4种Leptolyngbya、1种Chamaesiphon、1种Nostoc、1种Oscillatoria、2种Syne-chococcus和1种Chroococcidiopsis。同层不同位置土壤中蓝细菌种群亦有所不同,但每个取样点都有一些特有的蓝细菌种类。用常规方法对同一稻田土壤样品进行分离培养,根据蓝细菌鉴定图谱观察到类似Lyngbya、Oscillatori-a、Chroococcidiopsis及Nostoc的蓝细菌,但显微镜下无法准确分类。比较结果表明采用DGGE法比常规培养法能更准确进行蓝细菌多态性鉴定。 相似文献
13.
《Soil biology & biochemistry》2001,33(4-5):697-699
Fungal community analysis using 18S rDNA primer pairs and denaturing gradient gel electrophoresis of PCR products (Vainio, E.J., Hantula, J., 2000. Mycological Research 104, 927–936) was applied to field studies of the forest ecosystem. We report a DNA extraction method producing high quality DNA allowing successful PCR amplification from problematic samples without use of nested polymerase chain reaction (PCR) procedures. The analysis was found to be applicable for samples from environments of varying fungal diversities and high organic matter content: wood samples from fallen branches of trees, laboratory mini-ecosystems and forest humus samples. When the method was tested using replicate forest soil samples, it was shown to be highly reproducible. 相似文献
14.
通过一系列的对比实验发现,RNA电泳检测效果不仅与凝胶孔径大小和稀释与否相关,而且也与电泳时间长短和RNA上样量的多少有关。建立了一套可方便快捷地鉴定动物组织总RNA质量的方法,即在孔径宽度为7 mm的1.2% 琼脂糖凝胶上,仅需取3~5 μg 总RNA,加6×RNA上样缓冲液混合并用无RNase的灭菌双蒸水稀释到一定体积后上样,电泳15~30 min即可准确地鉴定RNA的质量。经实践证明,该方法实用性强,重复性好,具有良好的推广应用价值。 相似文献
15.
Achim Schmalenberger Christoph C. Tebbe Michael A. Kertesz Harold L. Drake Kirsten Küsel 《European Journal of Soil Biology》2008,44(5-6):495
Genetic fingerprinting methods such as denaturing gradient gel electrophoresis (DGGE) and single strand conformation polymorphism (SSCP) are only able to separate about 20–40 well-distinguishable bands (signals) within each sample. As a result, the diversity of 16S rRNA genes within biological samples may be underestimated, because multiple sequences can migrate at the same rate to form a single band. This study reports a two-dimensional SSCP fingerprinting method that has the capability to resolve hundreds of signals in a single fingerprint by using different gel temperatures in the two dimensions of the separation (20 °C and 30 °C, respectively). Unlike previous two-dimensional approaches, the method presented in this study does not rely on DNA products of variable lengths but is able to separate 16S rRNA gene fragments of the same length. To demonstrate the effectiveness of this new method, DNA samples from oxic and anoxic zones of an acidic fen were examined. Whereas one-dimensional SSCP fingerprints indicated high similarity (>93%) between 16S rRNA gene fragments from oxic and anoxic zones of the fen, the two-dimensional SSCP approach virtually found no similarities. 相似文献
16.
EMSA全称为电泳迁移率改变分析法，是目前研究核转录因子体外结合特性最常用的方法，但同时同位素对人体会有潜在的放射性损伤，使实验操作变得复杂。为了提高EMSA实验的稳定性和成功率，以降低实验人员接触同位素的频率，本研究对放射性EMSA实验体系进行了优化。结果表明：探针的纯化有效地降低了自显影的背景；蛋白的离心有利于获得整齐清晰的条带；采用350V的电压并且在4°C进行电泳，保证了电泳过程中DNA-蛋白复合物的稳定结合；拭去凝胶表面带有放射性的液体，并将凝胶附着于有一定硬度的纸片有利于得到清晰的显影图片。试验表明优化后的体系有效地提高了EMSA实验的稳定性和成功率。 相似文献
17.
Abha Jain A. K. Roy P. Kaushal D. R. Malaviya S. N. Zadoo 《Genetic Resources and Crop Evolution》2006,53(2):339-347
Isozyme banding pattern was studied in Guinea grass (Panicum maximum Jacq.), a widely cultivated grass having good fodder value. Similarity among 63 accessions collected from diverse sources
was worked out using five enzyme systems (SOD, GOT, ACP, Esterase and Peroxidase) following horizontal starch gel electrophoresis.
Biochemical markers such as isozymes are useful supplements in identifying the genetic variation present in any crop. A total
of 35 clear and unambiguous bands were used for analysis of which 8 bands were monomorphic. Polymorphism exhibited by 27 bands
from all five enzyme systems indicate presence of considerable diversity in this species. The dendrogram generated after UPGMA
and SAHN cluster analysis using Jaccard genetic distance showed that 63 accessions from diverse geographical locations could
be grouped in three main clusters, of which two could further be divided into sub-clusters. Although the clusters comprised
members from different locations, most of the accessions from similar geographical locations tended to cluster in same group. 相似文献
18.
Jun Murase Miho Shimizu Motoki Hayashi Kazuo Matsuya Makoto Kimura 《Soil Science and Plant Nutrition》2005,51(1):83-90
Percolating water was sampled from the plow layer and subsoil layer in a Japanese paddy field, and the bacterial communities were compared together with floodwater by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) targeting a partial 16S rRNA gene and subsequent sequencing. The number of DGGE bands ranged from 16 to 28 with no significant differences among the sampling sites and times. Only 2 bands were common for the three sources of water samples. DGGE bands specific for the floodwater samples and percolating water samples from the plow layer were identified, while percolating water samples from the subsoil layer did not show specific bands but displayed common bands to those of the floodwater samples (7 bands) and percolating water samples from the plow layer (1 band). Cluster analysis of the DGGE banding patterns showed a distinct clustering in the samples of percolating water from the plow layer and a closer relationship between the others. These results suggest that the bacterial communities in percolating water changed during downward movement through the plow layer and subsoil layer. Sequences of the DGGE bands specific for the samples of percolating water from the plow layer showed a close relationship with anaerobic bacteria such as iron-reducers or uncultured bacterial DNA isolated from environments that are considered to be less oxic. On the other hand, the sequences of the bands specific for the samples of floodwater and percolating water from the subsoil layer showed a close relationship with uncultured bacterial DNA isolated from freshwater environments. 相似文献
19.
The possibility of using electrophoresis to characterize varieties of pepper, Capsicum annuum and Capsicum frutescens cultivated in Nigeria was investigated. The SDS- polyacrylamide gel electropherogram of extracted total seed proteins of 10 breeding lines in each of the 6 varieties investigated, revealed a pattern in which 12 polypeptide bands with apparent molecular weight range of 22 to 98 kilodaltons could be distinguished. The result showed that the six varieties could be characterized on the basis of presence/absence and staining intensities of 7 polypeptide bands. It is suggested that SDS- polyacrylamide gel electrophoresis of seed proteins provides a useful analytical technique for the characterization of varieties of pepper and there may be genotype duplicates in the collection of Nigerian Capsicum germplasm. 相似文献