排序方式： 共有48条查询结果，搜索用时 31 毫秒
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靶位区域扩增多态性（target region amplification polymorphism,TRAP）是一种新型功能性分子标记,比较了L16（4^5）正交试验和单因素方法优化罗非鱼（Oreochromis spp．）TRAP反应体系。2种分析方法结果中dNTPs浓度、随机引物浓度、DNA浓度相同而镁离子（Mg2＋）浓度和TaqDNA聚合酶浓度不同,不同因素间存在一定的交互作用。正交设计方法比单因素法更简单、科学、合理。该试验获得罗非鱼15txLTRAP最优反应体系为DNA模板浓度为60ng、Mg2＋浓度为1．5mmol·L^-1、dNTPs浓度为0．3mmol·L^-1、TaqDNA聚合酶0．5U、随机引物浓度为7pmol·L^-1和固定引物浓度为10pmol·L^-1。该反应体系在4个罗非鱼群体中验证,表现出良好的稳定性和重复性,该反应体系的建立为TRAP标记应用于罗非鱼遗传多样性评价、种质鉴定、分子标记辅助育种等研究提供了新的手段。 相似文献
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为将新型分子标记-靶位区域扩增多态性(target region amplified polymorphism,TRAP)应用到遗传棉花多样性研究中,分析了不同浓度梯度的模板DNA、Mg2+ 、dNTPs、Taq酶与引物浓度对TRAP-PCR扩增结果的影响.确立了稳定性强、重复性好的棉花TRAP-PCR最佳反应体系:在15 μL的TRAP-PCR反应体系中,含10×buffer 1.5 μL,25 mM MgCl2 1.5 μL,2.5 mM dNTPs 2.0 μL,20 mM固定引物0.75 μL,20 mM随机引物0.15 μL,100 ng/μL DNA 0.5 μL,Tap酶0.2 μL.结果表明优化后的TRAP-PCR反应体系可应用于棉花遗传多样性分析研究中. 相似文献
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Kohji NAGAHORI Shigehisa IWAMOTO Akira MARUYAMA Shingo SUZUKI Kazuyoshi HOSOMICHI Takashi SHIINA Hiromi HARA Yutaka YOSHIDA Kei HANZAWA 《Animal Science Journal》2010,81(4):513-518
In the current study, we describe four novel members of the 90 kDa heat shock protein (HSP90) family expressed in Japanese quail, Coturnix japonica. The coding regions of the genes, CjHSP90AA1, CjHSP90AB1, CjHSP90B1 and CjTRAP1, exhibited more than 94% similarity to their related genes in chicken. The putative proteins encoded by these quail genes contained motifs considered essential for HSP90 gene function. In addition, the predicted proteins were more similar to HSP90AA1, HSP90AB1, HSP90B1 and TRAP1 proteins expressed in vertebrates than they were to other members of the HSP90 family. Exon numbers of CjHSP90AA1 (11), CjHSP90AB1 (12) or CjTRAP1 (18) are the same as the chicken and mammalian orthologs. Furthermore, gene order in the regions surrounding CjHSP90AB1 and CjTRAP1 has been preserved, providing evidence that the genomic regions were orthologous to HSP90‐containing regions in the chicken genome. The promoter regions of the genes also contained conserved motifs identified in related genes of chicken. However, the nucleotide sequences of the 5′‐flanking region of these genes were highly polymorphic. We also found that CjHSP90AA1 exhibited a robust response to heat shock treatment. Taken together, the data suggest that CjHSP90AA1, CjHSP90AB1, CjHSP90B1 and CjTRAP1 encode orthologs of HSP90AA1, HSP90AB1, HSP90B1 and TRAP1, respectively. 相似文献
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运用TRAP标记技术,选取10个多态性较好的引物组合对荷包红鲤、黄河鲤、建鲤、兴国红鲤和黑龙江野鲤等5个鲤群体进行遗传多样性分析,其中固定引物是根据目标候选基因-GHR基因的序列设计。结果表明,共扩增出168个位点,其中多态性位点134个,平均多态位点比例为80.41%,平均多态性信息含量为0.29。分子变异方差分析(AMOVA)结果显示群体内的方差贡献率达96.97%,表明各个鲤群体内存在较大的遗传变异。种群再分效应固定指数(FST=0.030 26,P＜0.05)表明不同鲤群体间有显著的遗传分化。基于目标候选基因的TRAP标记对5个鲤群体进行聚类分析,结果表明建鲤和兴国红鲤首先聚成一类,然后黄河鲤和黑龙江野鲤聚为一类,再与荷包红鲤聚为一类。 相似文献
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【目的】开发厚朴新型分子标记,分析广东乐昌龙山林场厚朴资源的亲缘关系,以方便后续进行
厚朴优株选择和优良品种选育等研究工作。【方法】以 14 个由广东乐昌龙山林场种植的、种源来自江西省各
地的药用植物厚朴为材料,根据 GenBank 中厚朴 cDNA 序列信息,利用 NCBI（https://www.ncbi.nlm.nih.gov）上
Primer BLAST 工具设计厚朴靶区域扩增多态性（Target Region Amplified Polymorphism, TRAP）标记固定引物,与
SRAP（Sequence Related Amplified Polymorphism）随机引物配对,扩增厚朴基因组 DNA,开发厚朴 TRAP 标记。
统计 TRAP 标记与厚朴 SSR（Simple Sequence Repeat）标记扩增产物的电泳带型图谱,用 NTSYSpc2.10e 软件进
行聚类分析,分析 14 个厚朴资源的遗传多样性及其亲缘关系。【结果】 从 14 条固定引物与 5 条随机引物配组
形成的 70 对引物中共开发出 7 个扩增产物电泳条带清晰、带型丰富、多态性好的厚朴 TRAP 标记;构建了厚朴
资源的亲缘关系树状图,结果表明龙山林场厚朴资源遗传相似系数在 0.75~0.88 之间,母株与对应子代之间遗传
相似系数并不是最高。【结论】厚朴群体内基因交流频繁,遗传多样性丰富,新开发的 TRAP 标记将为厚朴优
株鉴别、优良品种选育、分子标记辅助选择等后续研究工作提供便利。 相似文献
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Yuan XU Zhiheng DU Jiayu LIU Hang SU Fangyong NING Shiquan CUI Lijuan WANG Jianming LIU Chuanshuai REN Shengwei DI Xiujuan BAI 《Integrative zoology》2022,17(1):105-114
Identifying the mechanism for sex determination in amphibians is challenging. Very little is known about sex determination mechanisms of Rana dybowskii, a species of importance to evolutionary and conservation biology. We screened for sex-linked molecular markers in R. dybowskii in China using target region amplification polymorphism with 2 fixed primers against the sequences of Dmrt1. We found 2 male-linked molecular markers in R. dybowskii, which were 222 bp and 261 bp long. The detection rates of 222 bp marker in males form Xinglong, Huadian, and Dandong were 93.79%, 69.64%, and 13.64%, respectively, while the rate in females from Huadian was 27.50%. Besides, the detection rates of 261 bp marker in the above 3 regions were only observed in males at the rate of 93.79%, 87.50%, and 32.73%, respectively. The inheritance patterns of sex-linked molecular markers showed that the 2 sex-linked molecular markers were heterozygous. Compared to the XY-male parent, progeny from XX-pseudo-male parent possessed lower sex reversal ratio at the same rearing temperature, and the proportion of female froglets from an XX-pseudo-male parent was more than 95% at low rearing temperature (15°C). Our findings suggest that R. dybowskii displays male heterogamety, and the 2 sex-linked molecular markers may have a guiding significance for the protection and utilization of R. dybowskii. 相似文献