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  

Construction of a Full-length cDNA Library and Preliminary Analysis of Expressed Sequence Tags from Elite Half-pipe Snowboarding Athletes Lymphocyte

作　　者：张泽彪[1] 赵玉华[1] 周文婷[1] 汪宇峰[1] 赵晨琼[1] 关伟军[2] 朱志强[1] 

机构地区：[1]哈尔滨体育学院,哈尔滨150008 [2]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《首都体育学院学报》2017年第3期254-258,共5页Journal of Capital University of Physical Education and Sports

基　　金：黑龙江省青年科学基金项目(QC2013C020)

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。Objective： The aim of this study is to construct a cDNA library of blood from elite half-pipesnowboarders. And we can get a large number of ESTs （Expressed Sequence Tags） from it by sequencing thegenes. We can also use it for gene screening and function prediction. Methods： The cDNA library of blood fromelite half-pipe snowboarders was constructed by a series of experiments extracting and mixing Total RNA ofseveral elite ice-snow athletes, synthesizing double-stranded DNA, proteinase K digestion, Sfi 1 enzymedigestion, connecting with vector, packaging phage λ and amplification. Then 500 clones from the library wererandomly selected to be sequenced. Low-quality samples in the results of sequence were removed by Chromassoftware. The relevant information of ESTs was captured by matching gene sequences with BLAST of GenBank.Then the information of ESTs was presented to PANTHR and GO（Gene Ontology） bioinformatics database tomake a basic classification and analysis and to get some important genetic information in the cDNA library ofathletes. Results： The titer of cDNA library unamplified was 1.32×106pfu/ml, and the titer of cDNA libraryamplified was 1.65×1010pfu/ml .The recombinant rate of cDNA library amplified was 90.6%. We finally got 360sequences whose length were between 300bp and 110bp, and the average length was 606bp. 201 Geneinformation with annotation（BLAST）was distributed on 27 pathways（PANTHER） , including TXN, MDH1,ARL1, ARPC3, ACTG1 and many other genes which were separately distributed on pathways associated withmetabolism, Oxidative stress, or Cardiovascular system function. So we speculate that it may be related tophysical ability. GO was used to analysis and classify gene sequences, and then 143 Genes with functionalannotation in the database were confirmed to participate in different functions which mainly include cellular component, molecular function and biological process. Conclusions： This study has successfully constructedafull-length athletes＇ cDNA library which co

关 键 词：单板U型场地滑雪 外周血淋巴细胞 全长CDNA文库 表达序列标签 

分 类 号：G804.2[文化科学—运动人体科学]