【摘要】：構(gòu)建一個(gè)優(yōu)秀單板U型場地滑雪運(yùn)動(dòng)員血液的cDNA文庫,經(jīng)測序從中獲取大量的表達(dá)序列標(biāo)簽(Expressed Sequence Tags)ESTs,用于基因篩選和功能預(yù)測。方法:通過對多名優(yōu)秀冰雪運(yùn)動(dòng)員血液樣品的總RNA進(jìn)行提取、混合總RNA,合成雙鏈cDNA、蛋白酶K消化、Sfi1酶切并連接載體、λ噬菌體的包裝,經(jīng)擴(kuò)增最終獲得優(yōu)秀冰雪運(yùn)動(dòng)員cDNA文庫;隨后從文庫中隨機(jī)挑選500個(gè)克隆提交至北京諾賽基因組研究中心有限公司進(jìn)行測序,測序結(jié)果利用Chromas軟件去低質(zhì)量樣品,使用Gen Bank中的BLAST功能對所獲序列進(jìn)行比對獲取相關(guān)信息,將獲得的ESTs信息提交PAN THR和GO(Gene O ntology)生物信息數(shù)據(jù)庫進(jìn)行基本歸類分析,從中獲取運(yùn)動(dòng)員c DN A文庫中所涵蓋的重要遺傳信息。結(jié)果:得到滴度分別為1.32×106 pfu/m L和1.65×1010 pfu/m L的未擴(kuò)增文庫和擴(kuò)增后文庫,擴(kuò)增后文庫的重組率為90.6%;得到長度在300~1 100 bp序列360個(gè),平均長度606 bp;通過BLAST比對得出201條具有注釋的基因信息,利用PANTHER數(shù)據(jù)庫基本分析發(fā)現(xiàn)這些基因分布在27個(gè)pathway上,其中TXN、MDH1、AR L1、AR PC3、ACTG1等基因分別分布在參與新陳代謝、氧化應(yīng)激反應(yīng)、心血管系統(tǒng)機(jī)能等相關(guān)聯(lián)的信號通路上,推測其可能與運(yùn)動(dòng)能力存在關(guān)聯(lián)。通過GO分析對所測序列進(jìn)行歸類得到在該數(shù)據(jù)庫中具有功能注釋的基因143個(gè),參與31項(xiàng)不同功能,主要包括細(xì)胞組分、分子功能和生物過程。結(jié)論:構(gòu)建了一個(gè)全長的運(yùn)動(dòng)員cDNA文庫,其質(zhì)量符合構(gòu)建cDNA文庫的標(biāo)準(zhǔn)。
[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.
【作者單位】： 哈爾濱體育學(xué)院;中國農(nóng)業(yè)科學(xué)院北京畜牧獸醫(yī)研究所;
【基金】：黑龍江省青年科學(xué)基金項(xiàng)目(QC2013C020)
【分類號】：G863.1
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本文編號：2261156