共查询到20条相似文献，搜索用时 171 毫秒
1.
LTR反转录转座子是植物基因组中重要的组成部分,可作为遗传工具在生物学研究上发挥重要作用。为探究毛竹中具有潜在活性的LTR反转录转座子,解析LTR反转录转座子转座激活机制,本研究从毛竹基因组中克隆1条典型的LTR反转录转座子序列,命名为PHRE8,系统地分析了PHRE8的结构特征、插入特性和进化关系,并利用荧光定量PCR检测PHRE8在DNA甲基化抑制剂(5-氮杂胞苷)、辐射、高温、低温、高盐逆境胁迫下的转录水平。结果表明,PHRE8属于Ty3-gypsy超家族,全长 5 296 bp,具有完整的GAG与POL结构域,插入时间约为123.07万年,是具有理论潜在活性的转座子;在不同胁迫处理下,与野生实生苗相比,PHRE8的相对表达量均有所增加,猜测PHRE8在不同胁迫条件下,会激发其转录活性,影响宿主基因组结构和基因表达模式的变化,以适应外界环境改变。本研究结果为进一步探究毛竹基因组中活性转座子的功能奠定了一定的理论基础。 相似文献
2.
3.
LTR-逆转座子是构成基因组特别是植物基因组的重要组分。它们在寄主基因组的进化过程中起到重要作用。马铃薯是重要的经济作物和粮食作物,其全基因组序列的公布为进一步研究其遗传组成和演化提供了基础。本文以马铃薯全基因组序列为材料,用结构分析和同源比对的方法分离得到9318个完整的LTR_逆转座子,7281个非完整（trtmcated）LTR-逆转座子元件和3657个soloLTR元件。进一步研究表明,gypsy类转座子在距今两百万年（millionyearsago,MYA）时转座活性被抑制,而copia类元件活跃至今。马铃薯和番茄比较基因组学的研究表明,LTR-逆转座子序列变异率为18．73％,远高于基因序列的7．37％和CDS序列的5．01％。 相似文献
4.
硫代葡萄糖甙(glucosinolates,GS)是一种含氮、硫的重要植物次生代谢产物,是十字花科蔬菜风味的主要来源。本研究根据拟南芥(Arabidopsis thaliana)和大白菜(Brassica rapa ssp.pekinensis)基因组序列信息,比对GS生物合成相关基因的序列信息,发现大白菜中与GS含量相关的候选基因102个,分布于10条染色体上。拟南芥GS合成基因在大白菜中的同源基因个数分别为:零拷贝基因8个、单拷贝基因13个、2拷贝基因17个、2个以上多拷贝基因14个。通过FASTPCR6.0软件分析目标基因上下游序列各5kb,共开发出237个简单重复序列(SSR)位点,16个基因没有发现SSR位点,72个基因存在1~4个SSR位点,5个基因存在5个SSR位点,4个基因存在6个SSR位点,4个基因存在7个SSR位点,1个基因存在8个SSR位点。所开发的SSR位点中,共发现4种重复类型:单核苷酸重复最多,共122个,占SSR总数的51.4%;其次是二核苷酸重复类型,共48个,占SSR总数的20.3%;三核苷酸重复类型最少,共7个,占SSR总数的3.0%,另外60个SSR无明显的重复类型。具有SSR位点的86个基因中,77个基因可以设计SSR特异引物,其中49对特异引物在供试的75份大白菜高代自交系中得到有效扩增。其中16对引物具有多态性,占全部引物的20.8%;18对引物无多态性,占全部引物的23.4%;15对引物杂带多,占全部引物的19.5%;另外28对引物无扩增产物,占全部引物的36.4%。最终11对获得了清晰多态性扩增产物,共检测出26个等位基因。本研究为分子标记辅助选择培育高有益GS成分的大白菜新品种提供了基础资料,进而加快大白菜营养品质育种的进程。 相似文献
5.
家蚕胞质肌动蛋白基因启动子的克隆及piggyBac转座子表达载体的构建 总被引：4，自引：0，他引：4
摘要: 利用PCR方法从家蚕（Bombyx mori）总DNA中克隆到细胞质肌动蛋白基因（cytoplasmic actin）启动子片段，序列分析表明，该片段中含有典型的组成型表达的细胞质肌动蛋白基因A3（BmA3）调控序列：SRE元件（serum response element）和ActE1元件(active element 1)。该序列的核苷酸序列与来自欧洲的一个家蚕品种的序列同源性为94%。通过多次重组，将A3启动子片段(不含信号肽序列)与转座子 piggyBac的转座酶编码区融合构建成辅助质粒；将其(含信号肽序列和部分编码区)与报告基因多管水母绿色荧光蛋白基因gfp融合，插入到人工合成的转座子piggyBac两反向重复末端序列之间，进而构建成基于转座子piggyBac的转基因载体。研究中构建的转座子转基因载体仅6.4 kb，并在两反向重复末端序列之间设计了1个多克隆位点区，便于目的基因的插入。 相似文献
6.
玉米(Zea mays L.)病害的发生和流行是限制玉米高产稳产的重要因素。玉米细菌性褐斑病是目前具有潜在危害性的病害之一,在抗性基因遗传规律和染色体定位研究方面取得一定进展,但在基因克隆、功能鉴定等研究上进展较为缓慢,利用构建的Fosmid文库克隆抗病候选基因是目前较为快捷和有效的方法之一。本研究以玉米细菌性褐斑病的抗病自交系F349为材料构建了玉米Fosmid文库,克隆总数约为3.7×105个,平均插入片段长度为32 kb,实现4.73倍的玉米基因组覆盖,筛选到任意基因或序列的概率达到了99.12%。基于该文库,利用不同引物组合进行了两个类受体蛋白激酶基因(Psy1和Psy2)的克隆筛选,得到14个阳性克隆,其中4个阳性克隆包含Psy2基因和Psy1基因部分片段;3个阳性克隆包含完整Psy1基因;对其中两个阳性克隆13-12F-23和32-4A-6的测序结果显示,Psy1基因的启动子和内含子分别被插入了10.0和13.5 kb的外源片段;13.5 kb的插入片段是LTR家族的huck反转录转座子,10.0 kb的插入片段是含有helitron转座元件的复杂转座子;扩增到了Psy1基因的cDNA全长,说明这两种类型反转录转座子的插入并未使基因失活。本研究为玉米基因的克隆和功能分析提供了一种可行的研究途径。 相似文献
7.
屠云洁 《农业生物技术学报》2007,15(6)
本研究利用线粒体细胞色素C氧化酶I (cytochrome c oxidase ⅠCOⅠ)这一特定基因的特定区段来做DNA条形编码的基础，选择其中二段序列（Bar1 Bar2）设计2个引物对6个中国地方鸡种的线粒体DNA进行扩增，以对其进行种品鉴定。本研究选择的2段序列中，以Bar1序列648bp（712位点到1359位点）多态性最高，突变位点最多，为24个，品种间平均多态性为3.860%，6个鸡种在此序列中有各自不同的特异位点，NJ聚类分析6个品种基本被聚为不同的类别，与形态学分类基本一致，该COⅠ基因的Bar1序列用于这些品种鉴定是可行的；而Bar2序列进行序列多态性分析，由于其多态性低，大多品种没有其特异位点，NJ聚类分析6个品种基本被聚为不同的类别，与形态学分类基本一致，不利于品种鉴定。从本研究结果看，利用COⅠ基因的DNA条形编码（DNA Barcodes）进行不同鸡品种鉴定具有方便、快捷、经济、准确等优点，结合传统形态学鉴定手段将会揭示更深层次的遗传多样性和系统进化关系, 经过进一步改进是用于品种鉴定的优良工具。 相似文献
8.
piggyBac转座系统作为一种遗传修饰的工具,在很多生物体如哺乳动物、昆虫、酵母中已经得到了广泛应用.然而,目前piggyBac转座系统在绿藻一莱茵衣藻(Chlamydomonas reinhardti)中是否有活性很少有入研究.本研究构建含有一个拷贝或多个拷贝并且能够稳定表达piggyBac转座元件的莱茵衣藻藻株.然后在含有转座元件的藻株中瞬时表达普通的piggyBac转座酶和密码子优化后的piggyBac转座酶,通过限制性片段长度多态性(restriction fragment length polymorphism,RFLP)分析piggyBac转座元件的转座现象.转座酶转入莱茵衣藻细胞内一周和两周之后与转入之前的RFLP图谱对比,有很明显的差异.转入转座酶一周后的TR1藻株原有的1.5kb大小的片段消失,同时出现了3条新的片段:2.8、4.2和6.5 kb.两周之后,这3条片段又发生了变化,表明转座现象存在并且持续发生.转入密码子优化后转座酶一周后的TR3藻株中,1.5、3和4kb大小的片段没有发生改变,但是6.5 kb大小的片段消失了,与此同时新出现了一个2.8 kb大小的片段.两周之后,TR3的限制性片段多态性图谱并没有发生变化.为了检测转座酶是否成功转入细胞中,我们在新的接头序列两侧进行侧翼PCR,同时对PCR产物进行序列比对分析,结果显示,piggyBac转座子偶尔会在非常规性位点5’-TTT-3’和5’-ACGCAG-3’处发生剪切,而常规性剪切位点发生在5’-TTAA-3’处.研究结果表明,piggyBac转座酶对莱茵衣藻体内piggyBac转座子具有转座作用,piggyBac转座系统可以用于莱茵衣藻的遗传学修饰. 相似文献
9.
为了研究不同剂量重离子辐射对植物产生的表观遗传学效应规律,采用2种不同的高传能线密度和不同剂量(0、0.2、2Gy)的12C6+放射流辐照干燥的水稻种子,分别选取辐射水稻三叶期及分蘖期的叶片,通过甲基化敏感扩增多态性技术(MSAP)进行CCGG位点的甲基化状态分析。结果表明,重离子辐射,尤其是高剂量的辐射明显造成水稻DNA甲基化的多态性水平的变化,其中DNA去甲基化的多态性水平明显高于DNA甲基化的多态性水平,且DNA甲基化位点更倾向于CNG。此外,利用全基因组甲基化测序进一步验证了重离子辐射对水稻DNA甲基化的影响。本研究结果为探讨空间辐射引起水稻表观遗传变化的分子机制提供了重要的理论依据。 相似文献
10.
转座子诱变生防枯草芽胞杆菌B-916及抗真菌活性相关基因的克隆 总被引：2，自引：0，他引：2
摘要：生防枯草芽胞杆菌(Bacillus subtilis)B-916对水稻纹枯病有很好的防治效果。研究报道了采用转座子诱变技术从8 500个突变株中筛选到3株抗水稻纹枯病菌(Rhizoctonia solani )下降的突变株。PCR验证了转座子插入到了突变株基因组DNA中。虽然突变株相对B-916的生长速度,生物量和最终活菌数没有显著差异;但培养液对水稻纹枯病菌毒力和溶血性能显著下降。HPLC分析表明,B-916分泌的脂肽类抗生素与伊枯草素保留时间不同;突变株M-1、M-2和M-3相对B-916产生脂肽类抗生素发生显著变化。反向PCR克隆转座插入位点周边序列及测序结果表明,转座突变的基因与编码脂肽类化合物合成和调控相关的malonyl coenzyme A transacylase、lipopeptide synthetase B和sensor histidine kinase蛋白基因具有高度同源性。通过基因步行法实验已克隆到插入位点全长基因,分别命名为BacD,BacD和Degs,递交到GenBank,接受序列号为FJ192262和FJ194461。进一步进行了生物信息学分析发现,B-916 编码lipopeptide synthetase B基因与目前已报道的相应基因存在低同源性区域。 相似文献
11.
Xinhai Li Yiqin Huang Jiansheng Li Harold Corke 《Genetic Resources and Crop Evolution》2001,48(2):189-194
Choix, a plant in the tribe Maydeae of the grass family, has been cultivated in Asia for several thousand years. It is a potential gene resource for improvement of other cereal crops because of its nutritional value and tolerance to stress. Genetic variation and relationships among 21 Choix lachryma-jobi L. accessions were characterized by random amplified polymorphic DNA (RAPD) markers. A total of 205 DNA fragments across all materials were amplified with 31 random primers, averaging 6.61 per primer. Among amplified fragments, 115 showed polymorphism averaging 3.71 per primer. Of amplified markers, 56.1% were polymorphic, indicating considerable variation at the DNA level among these accessions. Some fragments were accession-specific. Pair-wise genetic similarity (GS) among 21 accessions ranged from 0.809 to 0.301. The 21 accessions clustered into two major groups. Three exotic Choix accessions clustered together. Three other Choix accessions, collected from Guangxi, China, clustered into a cohesive subgroup. Four wild types of Choix clustered into the same subgroup. These results indicated that the classification by RAPD data reflected the differences in geographic origins and evolution in Choix. 相似文献
12.
Shigeki Nakayama 《Genetic Resources and Crop Evolution》2012,59(7):1389-1396
Sugarcane, one of the most important tropical crops, belongs the genus Saccharum. This genus consists of six species, four cultivated and two wild. The domestication histories of the four cultivated Saccharum species is an interesting and important topic of study. Previous studies have categorized the four cultivated species into two groups, one consisting only of S. edule and the other comprising S. officinarum, S. sinense and S. barberi. All four species have inherited the genomic DNA of S. robustum, one of two wild relative species. Saccharum species have large genomes with complex structures, as evidenced by chromosomes with a high degree of polyploidy, alloploidy and aneuploidy. Miniature inverted-repeat transposable elements (MITEs) are class II (DNA) transposons that disperse throughout the plant genome. In this study, a Tourist family MITE sequence with 18/19-bp terminal inverted repeats and a 2-bp target site duplication was newly identified from genomic DNA of S. robustum. The abundant accumulation of this MITE sequence in the sugarcane genome enabled the application of inter-MITE polymorphism (IMP) analysis to Saccharum. IMP analysis revealed the genetic relationships among all six Saccharum species and the domestication histories of the four cultivated species. 相似文献
13.
利用ISSR标记技术分析了28份香蕉种质的遗传多态性。从100个ISSR引物中筛选出8个多态性引物,共扩增出55条DNA带,其中46条为多态性带,占83.6%,平均每个引物扩增的DNA带数为6.88条。依相似系数0.73的水平,将香蕉28个品种划分为6大类。其中云南BB（BB）和东莞高把大蕉（ABB）在相似系数为0.94时,二者的亲缘关系较近。Pisang Ceylan（AAB）和FHIA-18（AAAB）相似系数水平接近为1,表明二者亲缘关系最近。本研究为香蕉遗传关系的建立及品种鉴定提供理论基础。 相似文献
14.
Summary Genetic diversity and patterns of geographic variation among collections of Paspalum scrobiculatum (kodo millet) and P. polystachyum were studied using molecular markers generated through the random amplified polymorphic DNA (RAPD) method. A high level of polymorphism in RAPD markers was observed among the individual accessions, demonstrating the high genetic diversity of the crop. The markers obtained from the RAPD method were analyzed with the cluster analysis, principal coordinates and minimum spanning tree methods. Three major groups were resolved, one representing the African accessions, and two for the Indian accessions. The accessions of the north African kodo millet and P. polystachyum (considered conspecific with P. scrobiculatum) were quite distinct. The Australian kodo millet showed higher affinity to the African types. The study demonstrated that the RAPD technique can be applied to resolving degrees and patterns of genetic variation at the population and species levels, identifying cultivars, and defining gene pools of this crop. 相似文献
15.
Oz Barazani Azat Atayev Bakhtiyor Yakubov Vladimir Kostiukovsky Konstantin Popov Avi Golan-Goldhirsh 《Genetic Resources and Crop Evolution》2003,50(4):383-389
Seedlings of Pistacia vera L. developed from seeds of two separate populations in Turkmenistan, Kepele and Agachli, were evaluated for their growth potential and genetic polymorphism. Plant growth rate as well as random amplified polymorphic DNA (RAPD) analysis showed distinct differences between the two populations. In plant height growth rate, 17 Agachli accessions were 1.3 times higher on average than that of 10 accessions of Kepele (significant at p = 0.046) and 1.2 times higher for trunk diameter growth rate (p = 0.062). Cluster analysis divided most accessions into two main genetic groups according to their geographic origin. The Agachli group was further divided into two subgroups. One Kepele accession (K4), was genetically different from the rest and clustered on a separate outgroup. Two Agachli accessions (A12 and A17) were outside the two main populations clusters. Accessions K9 and K10 from Kepele were exceptional and were clustered in each of the two Agachli's subgroups, indicating a close genetic relationship between the two populations. In addition, high similarity values (0.58–1.00) and small genetic distances reflect plausible gene flow between Kepele and Agachli, which are 100 Km apart. Mantel test revealed significant relationship between the RAPD and the morphological traits matrices, pointing to the genetic basis for the measured differences in the growth rate. Growing the accessions on the same plot, under similar conditions enabled the evaluation of genotypic differences. The combination of morphological traits and molecular markers will further assist in preservation of genetic variability and cultivation of useful genotypes of P. vera L. 相似文献
16.
Summary The use of random amplified DNA fragments as genetic markers in Coffea was investigated. Arbitrary oligonucleotides were used as primers to amplify genomic DNA of different coffee accessions representing major Coffea species by polymerase chain reaction. Intraspecific variation was easily detected in C. canephora and C. liberica whereas the primers assayed failed to reveal polymorphism between C. arabica accessions. Extensive interspecific variation was observed. Genetic relationships between Coffea species are deduced from the degrees of similarity in amplified product profiles. Random amplified DNA markers appeared to be of high value for characterization, analysis and utilization of coffee genetic resources. 相似文献
17.
刘庆昌 《农业生物技术学报》2008,16(5)
以甘薯抗茎线虫病品种徐781和感茎线虫病品种徐薯18的杂交F1分离群体的186株单株为材料,利用分离群体分组分析法（BSA法）和AFLP技术,在抗感池中共筛选了800对AFLP引物,结果表明其中245对引物具有多态性。用这245对引物检测两亲本以及建池单株,发现引物组合E2M23和E33M20分别在抗病单株中扩增出1条在感病单株中未出现的特异条带,长度分别约为500 bp和200 bp,认为这2个AFLP标记与甘薯抗茎线虫病基因连锁,分别命名为E2M23500和E33M20200。根据这2个AFLP标记对F1代186个单株的扩增结果,经Mapmaker 3.0软件分析,发现这2个分子标记与抗茎线虫病基因位于同一连锁群并紧密连锁,它们与抗茎线虫病基因间的遗传距离分别为6.94 cM和11.1 cM。用这2个分子标记对10个中国甘薯主栽品种进行检测,所得结果与常规方法鉴定结果完全一致,表明2个分子标记可用于甘薯抗茎线虫病分子标记辅助育种。 相似文献
18.
Yogesh Kumar Soon Jae Kwon Clarice J. Coyne Jinguo Hu Michael A. Grusak Theodore J. Kisha Rebecca J. McGee Ashutosh Sarker 《Genetic Resources and Crop Evolution》2014,61(5):965-977
The effective utilization of crop diversity held in genebanks depends on our knowledge of useful traits and available markers associated with the target traits. Target region amplification polymorphism (TRAP) was used to evaluate the genetic diversity and underlying relationships among 263 accessions of chickpea landraces maintained by the USDA-ARS Western Regional Plant Introduction Station in Pullman, WA, USA. Two-hundred sixty-two TRAP markers were amplified by eight primer combinations. Altogether, 110 (42 %) markers were polymorphic, the other 152 (58 %) displayed no variation. These polymorphic markers revealed important differences among the accessions, with an estimated, mean pair-wise genetic distance of 25.82 %, ranging from 2.8 to 50.0 %. Genetic distance analysis divided the accessions into two major groups, with 113 and 150 accessions each, and substantial association between molecular diversity and geographic origin was evident. Bayesian analysis of population structure revealed two groups (K = 2) with evidence for six sub-groups. Additionally, the population structure of a subset of 110 lines was determined (K = 3) for testing marker-trait associations (MTAs). Phenotypic traits included the concentrations of protein and nine mineral elements in the seeds. Two MTAs were significant (p < 0.01) for concentrations of Ca and K, and three MTAs were significant for Cu and Ni concentrations. The results indicate that this population is useful for genome-wide association studies on other economic traits given the level of genetic diversity uncovered and the marker-trait associations in seed minerals discovered. 相似文献
19.
A polymerase chain reaction (PCR) based approach involving the directed amplification of minisatellite DNA region (DAMD-PCR)
was used to identify accession specific DNA markers and study genetic relationships between and within 15 accessions corresponding
to 11 species in genus Capsicum. A touch down PCR profile and unique chemical concentration of ingredients resulted in reproducible and reliable DNA amplifications.
The number of amplified products varied from 1 to 12 fragments depending on the template DNA and the primers. The DAMD-PCR
technique provided a total of 38 accession specific DNA markers (diagnostic DAMD-PCR) which can be utilized in accession identification,
preservation and genetic studies of Capsicum germplasm. Based on 1,292 polymorphic and monomorphic DNA markers directed with 22 minisatellite specific primers, accessions
were divided into four major groups, three of which corresponded to the three distinct Capsicum complexes. Capsicum chacoense was found to be the most distinct species. 相似文献
20.
Yu-Xia Yang Wei Wu You-Liang Zheng Li Chen Ren-Jian Liu Chun-Yan Huang 《Genetic Resources and Crop Evolution》2007,54(5):1043-1051
Genetic diversity and relationships among 48 safflower accessions were evaluated using 22 inter-simple sequence repeats (ISSR)
primers. A total of 429 bands were amplified, and 355 bands (about 82.7%) were polymorphic. Five to forty-one polymorphic
bands could be amplified by each primer, with an average of 16.1 polymorphic bands per primer. The results showed that the
polymorphism of the safflower germplasm was higher at the DNA level. All the 48 accessions could be distinguished by ISSR
markers and were divided into 9 groups based on ISSR GS by using UPGMA method. The genetic relationships among the accessions
from different continents were closer. Comparatively, the genetic diversity of the accessions originated from Asia was higher,
from Europe assembled. The results also showed that the genetic variation of accessions from Indian and Middle Eastern safflower
diversity centers were relatively higher. ISSR is an effective and promising marker system for detecting genetic diversity
among safflower and give some useful information on its phylogenic relationships. 相似文献