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1.
BT型细胞质雄性不育水稻及其三系的线粒体DNA研究 总被引:2,自引:0,他引:2
用RAPD技术对BT型水稻胞质雄性不育系秀A及其保持系秀B、恢复系湘晴以及杂种F1代的线粒体DNA进行了比较分析。结果表明不育系与其保持系间存在显著差异;不育系与其F1之间mtDNA也存在差异。在引物OPJ-08的扩增产物中,秀A扩增出一条分子量为800bp的多态性片段,在引物OPK-10的扩增产物中,杂种F1扩增出一条分子量为900bp的片段。把这两片段回收、克隆并制备探针,OPJ-08800的Southern杂交结果显示不育系与其F1杂交图谱存在多态性;OPK-10900的Suthern杂交结果显示不育系与其保持系同存在差异。推测这两片段与育性可能有一定的联系。 相似文献
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应用AFLP分析技术,比较了红莲型水稻雄性不育系和保持系线粒体DNA,找到一不育系特异条带TA1。以该条带DNA为探针与不育系、保持系以及F1杂种线粒体总RNA杂交,结果显示该片段黄化苗期三者均转录一个RNA分子,但转录产物分子量各不相同,说明三者该片段阅读框架不同。经测序,该片段长202bp,序列内含密码子ATG、ATT、AGA、AGG以及正向重复序列5’TGTAC3’、5’ATTATTTT3’和倒重复序列5’GGGAAACA3’。上述特点表明该片段可能是某一蛋白编码序列,并可能与红莲型水稻雄性不育性状形成有关。 相似文献
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雄牛特异的SRY同源序列的扩增和分析 总被引:6,自引:0,他引:6
利用人、兔、鼠SRY序列设计引物,应用PCR扩增牛的SRY序列,获得200bp的雄牛特异的扩增片段。克隆该扩增片段,获得重组质粒pCH21,进行序列分析,并与人、兔和鼠SRY的对应区域比较,具有高度同源性。用pCH21 DNA作探针与牛的基因组DNA酶切图谱杂交,显示了雄牛特异的I.7kb的杂交带。分析200bp的PCR扩增片段是牛的SRY基因片段。用同一对引物扩增人和山羊的DNA样品,也获得雄性特异的200bp的扩增片段。 相似文献
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非洲绿猴肾细胞株AGMr(HindⅢ)—1高度重复顺序的克隆及初步应用 总被引:2,自引:0,他引:2
陈宇光 《Virologica Sinica》1997,12(2):119-124
从非洲绿猴肾细胞株分离高度重复顺序AGMr(HindⅢ)-1基因,以pUC18质粒为载体构建重组质粒pUC18/AGMr(HindⅢ)-1,转化E.coliJM83。酶切分析、SouthernBlot分析和序列分析均证明克隆成功,但所用的第165代Vero细胞株与另一非洲绿猴肾BSC-1细胞株的序列相比较,172个碱基中有6个位点突变。用该重组质粒作探针,对Vero细胞培养的狂犬疫苗作SouthernBlot分析表明,未经纯化的半成品中残余细胞DNA长度约400~5000bp。斑点杂交分析表明,杂交特异性良好,残余细胞DNA最小检出量约4pg。提示AGMr(HindⅢ)-1高度重复顺序可特异地应用于Vero细胞残余DNA检测。 相似文献
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以来自Polima胞质甘蓝型油菜雄性不育源转育获得的不育白菜‘Bpol97-05A’和其回交亲本即保持系‘Bajh97-01B’为材料,利用cDNA扩增片段长度多态性(cDNA-AFLP)技术获得一条长约330 bp的特异片段P1708,RT-PCR验证确认该序列为不育白菜材料所特有,经测序和BLAST比对,发现该片段除54 bp的插入序列外,其余部分与大白菜和甘蓝叶绿体ndhJ-trnF基因之间的一段序列完全一致. 根据基因区域两端的保守部位设计引物,以Polima不育白菜DNA和可育甘蓝型油菜的DNA为模板,分别获得了长约1 900 bp的序列,比较序列发现:不育白菜与可育白菜、甘蓝型油菜的DNA序列存在一定差异,‘Bpol97-05A’中除多个位点发生变异外,另有108 bp的插入序列,该插入由2个长度为54 bp的重复序列组成,重复序列中除5′端3个碱基CTT外,其余部分均与trnF基因3′端51 bp完全相同. 相似文献
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牛β-酪蛋白5′端上游调控序列的克隆和序列分析 总被引:3,自引:0,他引:3
该文用PCR扩增了牛β-酪蛋白基因5′-端上游调控序列,并对其进行了克隆和序列分析。采集成年母牛肝,提取DNA。在牛β-酪蛋白基因外显子1和上游调控区内设计引物,扩增其上游调控序列。两条引物长均为19个核苷酸,引物间跨度为635bp。以牛肝DNA为模板,进行PCR扩增,扩增产物在2%琼脂糖凝胶上电泳,可见特异的目的条带。从凝胶中回收目的片段,克隆到pGEM-T载体中。重组质粒提取DNA,进行序列分析。测序结果与文献发表的类似序列相比,仅有4个碱基不同,同源性达99.4%。表明获得了牛β-酪蛋白基因5′-端上游调控序列的克隆。 相似文献
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将缺少编码信号肽序列的人白细胞介素-11(hIL-11)546核苷酸cDNA,重组于质粒pBacPAK8构建重组转移载体pBacIL-11,与经线性化修饰的家蚕核型多角体病毒(BmBacPAK)DNA共转染家蚕培养细胞株BmN,获得了插入hIL-11基因的重组病毒。Southern杂交表明重组病毒基因组中含有hIL-11基因片段,RNA斑点杂交表明hIL-11基因得到了转录。重组病毒感BmN细胞株、家蚕幼虫和蛹,在细胞培养上清、细胞抽提物、幼虫和蛹的体液样品中,SDS-PAGE电泳分析都能检测得到表达产物的特异性条带;采用IL-11依赖细胞株B9-11和MTT法测定表达产物的生物活性,表明rIL-11基因分别在培养细胞和蚕体内得到了高效表达。 相似文献
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以来自Polima胞质甘蓝型油菜雄性不育源转育获得的不育白菜'Bpol97-05A'和其回交亲本即保持系'Bajh97-01B'为材料,利用cDNA扩增片段长度多态性(cDNA-AFLP)技术获得一条长约330bp的特异片段P1708,RT-PCR验证确认该序列为不育白菜材料所特有,经测序和BLAST比对,发现该片段除54bp的插入序列外,其余部分与大白菜和甘蓝叶绿体ndhJ-trnF基因之间的一段序列完全一致.根据基因区域两端的保守部位设计引物,以Polima不育白菜DNA和可育甘蓝型油菜的DNA为模板,分别获得了长约1900bp的序列,比较序列发现:不育白菜与可育白菜、甘蓝型油菜的DNA序列存在一定差异,'Bpol97-05A'中除多个位点发生变异外,另有108bp的插入序列,该插入由2个长度为54bp的重复序列组成,重复序列中除5′端3个碱基CTT外,其余部分均与trnF基因3′端51bp完全相同. 相似文献
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睾丸决定因子基因(Testis-determining factor,TDF)位于Y染色体短臂上,它的表达产物诱导睾丸组织的发生。SRY基因(Sex-determining Region of the Y)位于Y染色体的性别决定区内,许多特征显示SRY就是TDF。我们选用与SRY基因相应的引物,用PCR技术对正常人男女各10例的基因组DNA进行扩增。将特异扩增的男性SRY基因片段重组到质粒PUC12中,得到含有中国人SRY基因片段的克隆,命名为PSY-1、PSY-2。用[~(32)p]标记重组质粒中的SRY基因片段作探针,与PCR结果进行Southern杂交,男性样品均显示特异杂交带,女性样品为阴性。用末端终止法测定克隆的SRY基因片段的全部核苷酸序列为299bp,含有SRY基因中高度保守及功能特异性区域的240bp,我们对此进行了讨论。 相似文献
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Hadi Ravan Soheila Kashanian Nima Sanadgol Arastoo Badoei-Dalfard Zahra Karami 《Analytical biochemistry》2014
Specific and predictable hybridization of the polynucleotide sequences to their complementary counterparts plays a fundamental role in the rational design of new nucleic acid nanodevices. Generally, nucleic acid hybridization can be performed using two major strategies, namely hybridization of DNA or RNA targets to surface-tethered oligonucleotide probes (solid-phase hybridization) and hybridization of the target nucleic acids to randomly distributed probes in solution (solution-phase hybridization). Investigations into thermodynamic and kinetic parameters of these two strategies showed that hybridization on surfaces is less favorable than that of the same sequence in solution. Indeed, the efficiency of DNA hybridization on surfaces suffers from three constraints: (1) electrostatic repulsion between DNA strands on the surface, (2) steric hindrance between tethered DNA probes, and (3) nonspecific adsorption of the attached oligonucleotides to the solid surface. During recent years, several strategies have been developed to overcome the problems associated with DNA hybridization on surfaces. Optimizing the probe surface density, application of a linker between the solid surface and the DNA-recognizing sequence, optimizing the pH of DNA hybridization solutions, application of thiol reagents, and incorporation of a polyadenine block into the terminal end of the recognizing sequence are among the most important strategies for enhancing DNA hybridization on surfaces. 相似文献
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William J. Boecklen 《Ecology and evolution》2017,7(24):10486-10491
Syngameons are sets of species linked by interspecific hybridization. Common observations regarding the structure of syngameons are that hybridization propensity is not uniform across species and that patterns of hybridization are dominated by a few species. I use computer simulations to test these claims in naturally occurring syngameons selected from the literature and from personal observation. Natural syngameons, especially those involving plants, typically exhibit nonrandom structure: The first three order statistics for the number of hybrid partners and the variance in the number of hybrid partners are larger than chance alone would predict. The structure of two insect syngameons examined is not significantly different from random. To test a hypothesis that variation in hybridization propensity across species in natural syngameons is simply an artifact of hybridization opportunity, I examine the structure of four artificial syngameons (fertility relationships) produced by full diallel crosses. Three of four artificial syngameons exhibit nonrandom structure, as the observed variation in number of successful crosses is larger than chance alone would predict. In general, there are no significant results involving the order statistics. Finally, I discuss biogeographic, ecological, and phylogenetic hypotheses for variation in hybridization propensity across species in natural syngameons. 相似文献
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Hybridization of nucleic acids immobilized on solid supports 总被引:252,自引:0,他引:252
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Some technical tips are described on how to improve the discrimination between perfect and imperfect duplexes formed by hybridization
of fluorescently labeled oligonucleotides to biological microchips. Model experiments were performed to assess the precision
of the method. Effects of labeling on the efficiency of hybridization and some properties of competitive hybridization were
studied using short synthetic oligonucleotides and three most popular fluorochromes as examples. 相似文献
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秀丽隐杆线虫(Caenorhabditis elegans)是一种重要的模式生物,目前已经被广泛应用到生物对各种驱虫剂抗性机制的研究中。福尔马林被普遍用于鱼类寄生虫病的防治中,但是由于长期的应用,许多寄生虫对它产生了一定的抗药性。研究以秀丽隐杆线虫为研究对象,分析了它在福尔马林刺激下基因表达的改变情况。结果显示,经福尔马林处理后差异表达的有676个克隆,通过斑点杂交技术进一步筛选,对其中差异显著的161个克隆进行了测序分析。测序结果经BLAST分析发现:(1)细胞凋亡相关基因的表达发生上调,这些基因编码包括线粒体呼吸链相关蛋白、含TPR序列的蛋白SGT-1、热休克蛋白、氧化应激相关蛋白、胞吞过程相关蛋白、DNA复制和修复相关蛋白以及其他一些重要的凋亡相关蛋白;(2)编码重要的转录调节因子和信号转导相关蛋白如转录激活因子FosB/c-Fos、G蛋白、细胞周期蛋白B、钙结合蛋白、核小体装配蛋白NAP-1等的基因的表达发生上调;(3)能量代谢和蛋白质、脂肪、氨基酸代谢途径中的有些基因的表达也发生上调;(4)除了以上已知功能的基因外,还有一些未命名蛋白质基因。这说明福尔马林对秀丽隐杆线虫的影响除了诱导细胞凋亡之外,还影响其他细胞代谢活动的改变。实验为进一步研究生物体对福尔马林的抗性机制奠定了一定的基础。
相似文献
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本文对报春花属植物引种的历史和现状、育种途径和育种成果进行了综述.报春花属是报春花科最大的属,全世界约500余种,我国有296种.国外在杂交育种、多倍体育种、组织培养和体细胞融合等方面取得了许多研究成果,而国内有关报春花属植物育种的研究相对较少.目前通过杂交育种或多倍体育种等手段已培育出众多花色丰富、花型各异或不含报春碱的报春花新品种.今后我国应在保护种质资源的基础上,加强报春花新品种的培育,并尽快实现产业化生产. 相似文献
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不吸水链霉菌基因文库的构建及初步筛选 总被引:1,自引:0,他引:1
采用碱裂解法从不吸水链霉菌提取染色体DNA,用Sau3AI对染色体DNA进行部分水解,利用透析袋法回收3.5~9kb之间的DNA片段。以pUC18为载体,用BamHI对其进行酶切,再用热敏磷酸酶去磷酸化,酶切产物与外源DNA按一定比例混合后,加入T4DNA连接酶进行连接。连接产物用受体菌E.coliDH5α进行转化,根据α-互补性质产生的颜色反应,挑选白色菌落,构建了相应的不吸水链霉菌基因文库。分别利用特异性探针2-1-1及1-2-1对不吸水链霉菌基因文库进行筛选,利用DIG-Ⅱ-dUTP对特异性探针进行标记,与基因文库中的阳性转化子进行Southern杂交,通过显色反应对杂交结果进行检测,由于时间关系,暂未筛选出阳性克隆,但这些工作对以后的相关实验研究具有很好的借鉴作用。 相似文献
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Acute and Persistent Suppression of Preproenkephalin mRNA Expression in the Striatum Following Developmental Hypoxic-Ischemic Injury 总被引:1,自引:0,他引:1
Robert E. Burke Steven O. Franklin Charles E. Inturrisi 《Journal of neurochemistry》1994,62(5):1878-1886
Abstract: The striatum is vulnerable to hypoxic-ischemic injury during development. In a rodent model of perinatal hypoxia-ischemia, it has been shown that striatal neurons are not uniformly vulnerable. Cholinergic neurons and NADPH-diaphorase-positive neurons are relatively spared. However, it is unknown what classes of striatal neurons are relatively sensitive. One of the major classes of striatal neurons uses enkephalin as a neurotransmitter. We have studied the effect of early hypoxic-ischemic injury on this class of neurons using a quantitative solution hybridization assay for preproenkephalin mRNA in conjunction with in situ hybridization. Hypoxia-ischemia results in an early (up to 24 h) decrease in striatal preproenkephalin mRNA, which is shown by in situ hybridization to occur mainly in the dorsal portion of the striatum. By 14 days, whole striatal preproenkephalin mRNA and total enkephalin-containing peptide levels are normal. However, at 14 days, in situ hybridization reveals that regions of complete preproenkephalin mRNA-positive neuron loss remain in the dorsal region. Normal whole striatal levels are due to an up-regulation of preproenkephalin mRNA expression in the ventrolateral region of the injured striatum. Given the important role that the enkephalin-containing striatal efferent projection plays in regulating motor function, its relative loss may be important in the chronic disturbances of motor control observed in brain injury due to developmental hypoxic-ischemic injury. 相似文献