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在构建内蒙古阿尔巴斯白绒山羊次级毛囊兴盛期皮肤组织cDNA文库的基础上,随机挑取636个克隆从5’端开始测序,对序列进行特征分析。分析结果表明有41个ESTs与绵羊KAP6-1基因同源性大于95%,且期望值小于1e-10,进一步分析可被分为6个Qusters。从每个Cluster中挑取一个全长cDNA序列,其核苷酸序列和预测编码的蛋白质序列表明AY310753与绵羊KAP6-1最为相似、AY310750差别最大,AY310751和AY310752在编码区分别缺少36个的核苷酸或12个的氨基酸。 相似文献
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T. C. Fischer J. T. Klattig A. Gierl 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2001,103(6-7):1014-1021
The investigation of plant cytochrome P450 genes and enzymes is a field of growing interest. Apparently, an even greater diversity
of cytochrome P450 genes exists in plants in comparison to other eukaryotes. This may be due to their role in the biosynthesis
of secondary metabolites that are present in plants in an enormous variety. Most cloning approaches are hampered by the large
sequence diversity of plant cytochrome P450 genes. We present a method to clone divergent cytochrome P450 ESTs by a nested
RT-PCR-strategy. These ESTs were used for the subsequent cloning of the corresponding full-size cDNAs of divergent families
via cDNA-library screening. Sixteen cytochrome P450 genes belonging to different cytochrome P450-families have been identified
in this way, proving the efficacy of the strategy.
Received: 1 December 2000 / Accepted: 26 February 2001 相似文献
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Xiaoqian Xu Shuhui Song Qun Wang Fen Qin Kan Liu Xiaowei Zhang Songnian Hu Yunlong Zhao 《Molecular genetics and genomics : MGG》2009,282(2):197-203
The water flea Daphnia carinata (D. carinata) reproduces both sexually and parthenogenetically, yet little is known about the genes involved in these processes. To further
clarify the reproductive biology of Daphnia and elucidate their unique mechanism of reproductive transformation, we have generated and characterized an expressed sequence
tag (EST) data set from D. carinata. A set of 1,495 clusters were generated from sequencing 3,072 randomly chosen clones from a parthenogenetic, juvenile water
flea cDNA library. The nucleic acid and deduced amino acid sequences were compared with known GenBank sequences. Functional
annotation found that 959 clusters showed significant homology with known genes involved in a broad range of activities, including
metabolism, translation, development and reproduction, as well as genes involved in sensing environmental factors. We speculate
that genes involved in development and reproduction, along with genes that allow the organism to sense changes in the environment,
play important roles in the process of parthenogenetic reproduction and could be markers of the early steps of sexual differentiation.
Additionally, 86% of the D. Carinata unique sequences could be stringently mapped to the D. pulex genome, of which 125 mapped to intergenic and intronic regions on the current assembly. Our results provide practical insight
into crustacean reproductive biology, in addition to establishing a new animal model for reproductive and developmental biology.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Xiaoqian Xu and Shuhui Song contributed equally to this work.
Nucleotide sequence data reported are available in the GenBank databases under the accession numbers: GD269049−GD272045. 相似文献
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Atul Grover Maya Kumari Sadhana Singh Shivender Singh Rathode Sanjay Mohan Gupta Pankaj Pandey Sween Gilotra Devender Kumar Mohommad Arif Zakwan Ahmed 《Physiology and Molecular Biology of Plants》2014,20(1):139-142
Oil-rich seeds of Jatropha curcas are being focussed as a source of bio-diesel. However, prior to its industrial use, a lot of crop improvement efforts are required in Jatropha. Availability of a large number of EST sequences of Jatropha in public domain allow identification of candidate genes for several agronomic characters including oil content in seeds. Here, we have analysed 42,477 ESTs of Jatropha spanning 22.9 Mbp for microsatellites and fatty acid metabolism related sequences. Unigene sequences were built using CAP 3 programme resulted in 12,358 contigs and 5,730 singlets. Nearly, 8 % unigenes showed presence of microsatellites, slightly over-represented compared to their occurrence in ESTs. Most of the microsatellites were either di- or tri-nucleotide repeats, while other categories of tetra-, penta- and hexa-nucleotide repeats together constituted ~4 % of total microsatellites. Assessment of functional relevance of unigenes was carried out using Blast2GO using its default settings. The overall sequence similarity level against sequences in ‘nr’ database was >80 %. A total of 931 sequences that participated in any of the pathways related to fatty acid or lipid metabolism were found at GO level 6. Among these, GO terms “Fatty acid metabolic process” and “Fatty acid biosynthetic process” were most over-represented. Overall, our work has due relevance in identifying molecular markers for the candidate genes for oil content in Jatropha seeds, and will prove to be an important reference for further studies for identification of trait specific markers in Jatropha. 相似文献
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香猪肌肉组织cDNA文库的构建及其EST测序成功率的分析 总被引:9,自引:0,他引:9
以香猪背最长肌为实验材料,以Lambda ZAP II为载体,构建了肌肉组织的cDNA文库。结果表明,cDNA文库的滴度在3.4×107 pfu/ml以上,重组率在94%以上,插入片段大小平均在1.5kb以上。同时指出,如果从3′端测序,多于30 个碱基T的插入片段是造成ESTs测序成功率低的主要因素。
Abstract:Using Longissimus Dorsi muscle as material and Lambda ZAP II as Vector,Xiang Pig Longissimus Dorsi muscle cDNA library has been constructed in our study.The results showed that the tritation of the library was 3.4×107 pfu/ml,the recombinant percentage was 94%,and the fragment length of inserted average cDNA were 1.5kb.The study pointed out that the more than 30 T insertion is the major factor for low percetage if sequencing the 3′-end. 相似文献
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47个早期人胚胎低丰度表达基因ESTs筛选及结果分析 总被引:1,自引:0,他引:1
构建高质量cDNA文库在基因克隆、mRNA差异展示、表达序列标签测序和基因定位等研究中具有十分重要的作用。为了从早期胚胎中分离人类新基因,构建了受精后3周龄的人cDNA文库,用标记的一链cDNA探针对该文库的6508个克隆子进行菌落原位杂交,得到1677个无任何杂交信号的低丰度表达克隆子,从中随机挑选了47个进行5′端部分测序,将测序结果与三大基因库进行序列同源性比较,发现18个克隆(38.3%) 相似文献
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