摘要
【目的】寻找偃松种子萌发过程的内参基因,为研究偃松种子萌发过程提供必要的分子生物学基础,丰富偃松基因资源。【方法】利用偃松种子萌发RNA-Seq数据和Genbank数据库对比筛选偃松EF-1α基因,通过生物信息网站及软件对其进行生物信息学分析。【结果】得到一条偃松EF-1α基因,命名为PpEF-1α。该基因具有完整的开放读码框(ORF),ORF长1344 bp,编码447个氨基酸,其氨基酸序列与挪威云杉EF-1α氨基酸序列(GenBankID:CAC27139.1)相似度最高,可达97.46%。PpEF-1α基因存在一个PTZ00141超家族位点。该基因编码的蛋白为亲水性稳定蛋白,具有α螺旋、延伸链以及无规则卷曲二级结构。根据实验室前期转录组分析结果显示,PpEF-1α基因在此过程中稳定表达。【结论】找到偃松种子萌发过程中的一个内参基因PpEF-1α,但其表达趋势有待于进一步的实验验证。
【Objective】In order to provide a necessary molecular biological basis for the study of seed germination process of Pinus pumila,and enrich the gene resources of Pinus pumila,we need to identify a reference gene for RT-PCR analysis.【Method】Screening EF-1αgene of Pinus pumila base on the comparison of RNA-Seq data of seed germination of Pinus pumila and genbank database,and analyzed the gene through biological information website and software.【Result】An EF-1αgene of Pinus pumila was screened,named PpEF-1α.The gene contained complete ORF(opening reading frame),ORF was 1344 bp,encoding 447 amino acids,the amino acid sequence had the highest similarity with the EF-1αprotein of Picea abies(GenBankID:CAC27139.1),up to 97.46%.There was a PTZ00141 superfamily loci in PpEF-1αgene.The protein encoded by PpEF-1αis a hydrophilic and stable protein,had alpha help,extended strand and random coil secondary structure.According to the results of transcripts in the early stage of the laboratory,it showed that the expression of PpEF-1αgene was stable in the germination process of Pinus pumila seed.【Conclusion】A reference gene for RT-PCR analysis during seed germination of Pinus pumila was finded,named PpEF-1α.However,the expression trend of it needs to be further verified by experiments.
作者
王思瑶
于宏影
丛日征
闫晓娜
周鑫
WANG Si-yao;YU Hong-ying;CONG Ri-zheng;YAN Xiao-na;ZHOU Xin(Harbin Research Institute of Forestry Machinery,National Forestry and Grassland Administration,Harbin 150086;China Research Center of Cold Temperate Forestry,CAF,Harbin 150086,China)
出处
《温带林业研究》
2019年第4期47-53,共7页
Journal of Temperate Forestry Research
基金
林木遗传育种国家重点实验室开放基金(K2019105)
中央级公益性科研院所基本科研业务费专项资金(CAFYBB2016ZD009、CAFYBB2017ZX001-6)
