优秀单板U型场地滑雪运动员外周血淋巴细胞cDNA文库构建及ESTs分析
发布时间：2018-10-10 07:26
【摘要】：构建一个优秀单板U型场地滑雪运动员血液的cDNA文库,经测序从中获取大量的表达序列标签(Expressed Sequence Tags)ESTs,用于基因筛选和功能预测。方法:通过对多名优秀冰雪运动员血液样品的总RNA进行提取、混合总RNA,合成双链cDNA、蛋白酶K消化、Sfi1酶切并连接载体、λ噬菌体的包装,经扩增最终获得优秀冰雪运动员cDNA文库;随后从文库中随机挑选500个克隆提交至北京诺赛基因组研究中心有限公司进行测序,测序结果利用Chromas软件去低质量样品,使用Gen Bank中的BLAST功能对所获序列进行比对获取相关信息,将获得的ESTs信息提交PAN THR和GO(Gene O ntology)生物信息数据库进行基本归类分析,从中获取运动员c DN A文库中所涵盖的重要遗传信息。结果:得到滴度分别为1.32×106 pfu/m L和1.65×1010 pfu/m L的未扩增文库和扩增后文库,扩增后文库的重组率为90.6%;得到长度在300~1 100 bp序列360个,平均长度606 bp;通过BLAST比对得出201条具有注释的基因信息,利用PANTHER数据库基本分析发现这些基因分布在27个pathway上,其中TXN、MDH1、AR L1、AR PC3、ACTG1等基因分别分布在参与新陈代谢、氧化应激反应、心血管系统机能等相关联的信号通路上,推测其可能与运动能力存在关联。通过GO分析对所测序列进行归类得到在该数据库中具有功能注释的基因143个,参与31项不同功能,主要包括细胞组分、分子功能和生物过程。结论:构建了一个全长的运动员cDNA文库,其质量符合构建cDNA文库的标准。
[Abstract]:A cDNA library of the blood of an excellent snowboarder was constructed and a large number of expressed sequence tags (Expressed Sequence Tags) ESTs,) were obtained by sequencing for gene screening and functional prediction. Methods: by extracting the total RNA from the blood samples of many excellent ice and snow athletes, mixing the total RNA, and synthesizing double strand cDNA, protease K to digest the Sfi1 vector, the 位 phage was packaged, and the cDNA library of the excellent ice and snow athletes was obtained. Subsequently, 500 clones were randomly selected from the library and submitted to Beijing Norse Genome Research Center Co., Ltd. for sequencing. The sequencing results were used to remove low-quality samples using Chromas software. The BLAST function of Gen Bank is used to compare the obtained sequences to obtain the relevant information, and the obtained ESTs information is submitted to the PAN THR and GO (Gene O ntology) biological information databases for basic classification and analysis. The important genetic information contained in the athlete's c DN A library is obtained from this paper. Results: the unamplified and amplified libraries with titer of 1.32 脳 10 ~ 6 pfu/m / L and 1. 65 脳 1010 pfu/m / L were obtained, respectively. The average length of 606 bp; was compared with BLAST, and 201 annotated genes were obtained. Using the PANTHER database, we found that these genes were distributed on 27 pathway, among which TXN,MDH1,AR L1, AR PC3,ACTG1 and other genes were involved in metabolism and oxidative stress respectively. In the signal pathways associated with cardiovascular system function, it is speculated that it may be associated with motor ability. 143 genes with functional annotation in the database were classified by GO analysis and involved in 31 different functions, including cellular components, molecular functions and biological processes. Conclusion: a full-length athlete cDNA library is constructed, and its quality accords with the standard of constructing cDNA library.
【作者单位】： 哈尔滨体育学院;中国农业科学院北京畜牧兽医研究所;
【基金】：黑龙江省青年科学基金项目(QC2013C020)
【分类号】：G863.1
，
本文编号：2261156
[Abstract]:A cDNA library of the blood of an excellent snowboarder was constructed and a large number of expressed sequence tags (Expressed Sequence Tags) ESTs,) were obtained by sequencing for gene screening and functional prediction. Methods: by extracting the total RNA from the blood samples of many excellent ice and snow athletes, mixing the total RNA, and synthesizing double strand cDNA, protease K to digest the Sfi1 vector, the 位 phage was packaged, and the cDNA library of the excellent ice and snow athletes was obtained. Subsequently, 500 clones were randomly selected from the library and submitted to Beijing Norse Genome Research Center Co., Ltd. for sequencing. The sequencing results were used to remove low-quality samples using Chromas software. The BLAST function of Gen Bank is used to compare the obtained sequences to obtain the relevant information, and the obtained ESTs information is submitted to the PAN THR and GO (Gene O ntology) biological information databases for basic classification and analysis. The important genetic information contained in the athlete's c DN A library is obtained from this paper. Results: the unamplified and amplified libraries with titer of 1.32 脳 10 ~ 6 pfu/m / L and 1. 65 脳 1010 pfu/m / L were obtained, respectively. The average length of 606 bp; was compared with BLAST, and 201 annotated genes were obtained. Using the PANTHER database, we found that these genes were distributed on 27 pathway, among which TXN,MDH1,AR L1, AR PC3,ACTG1 and other genes were involved in metabolism and oxidative stress respectively. In the signal pathways associated with cardiovascular system function, it is speculated that it may be associated with motor ability. 143 genes with functional annotation in the database were classified by GO analysis and involved in 31 different functions, including cellular components, molecular functions and biological processes. Conclusion: a full-length athlete cDNA library is constructed, and its quality accords with the standard of constructing cDNA library.
【作者单位】： 哈尔滨体育学院;中国农业科学院北京畜牧兽医研究所;
【基金】：黑龙江省青年科学基金项目(QC2013C020)
【分类号】：G863.1
，
本文编号：2261156
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