偃松PpSS和PpSE基因克隆及序列分析  

Cloning and Sequence Analysis of PpSS and PpSE Genes in Pinus pumila

作　　者：王思瑶 于宏影 刘洋[1] 李翊宁 周鑫 南方 WANG Si-yao;YU Hong-ying;LIU Yang;LI Yi-ning;ZHOU Xin;NAN Fang(Harbin Research Institute of Forestry Machinery,National Forestry and Grassland Administration,Harbin 150086;Research Center of Cold Temperate Forestry,CAF,Harbin 150086)

机构地区：[1]国家林业和草原局哈尔滨林业机械研究所,哈尔滨150086 [2]中国林业科学研究院寒温带林业研究中心,哈尔滨150086

出　　处：《温带林业研究》2023年第1期35-40,共6页Journal of Temperate Forestry Research

基　　金：新兴食用油料树种种质资源调查收集(2019FY100802-06)。

摘　　要：【目的】克隆偃松甾醇代谢途径关键基因SS和SE,为促进偃松甾醇高效合成奠定基础,为药用偃松遗传改良提供重要基因资源。【方法】利用偃松RNA-Seq数据筛选偃松SS和SE基因并克隆,通过生物信息网站及软件对其进行序列分析。【结果】从偃松中克隆2个甾醇代谢途径关键基因SS和SE,分别属于SS和SE基因家族,命名为PpSS和PpSE。二者均具有完整的开放读码框(ORF),分别长1230bp和1647bp,编码409和548个氨基酸。PpSS编码的氨基酸序列与马尾松SS家族蛋白(GenBankID:AHI96421.11)相似度最高,为95.11%,PpSE编码的氨基酸序列与银杏SE家族蛋白(GenBankID:AVG44184.1)相似度最高,为80.98%。PpSS蛋白存一个异戊二烯合成酶C1超家族位点,PpSE蛋白存在一个鲨烯单加氧酶超家族位点。【结论】PpSS和PpSE分别为偃松SS和SE基因家族中的新基因,两个基因的获得为进一步挖掘其在偃松甾醇代谢中的调控机制研究奠定了基础。【Objective】In order to provide technical support and theoretical basis for the genetic transformation of Pinus pumila sterols metabolic engineering,the key genes SS and SE in the sterol metabolic pathway of Pinus pumila were cloned.【Method】Screening and clone SS and SE genes of Pinus pumila base on the RNA-Seq data of Pinus pumila,then analyzed the genes through biological information website and software.【Result】SS and SE genes from Pinus pumila in response to the sterol metabolic pathway were cloned,named PpSS and PpSE,and the genes belong to SS and SE gene family,respectively.The two genes both contain complete ORF(opening reading frame),PpSS gene ORF was 1230 bp,encoding 409 amino acids,PpSE gene ORF was 1647 bp,encoding 548 amino acids.The amino acid sequences were the most similar to SS and SE family protein of Pinus massoniana(GenBankID:AHI96421.11)and Ginkgo biloba(GenBankID:AVG44184.1),respectively,the similarity were up to 95.11%and 80.98%.There was one isoprenoid biosyntheses enzymes class 1 in PpSS protein,and one squalene monooxygenase super family in PpSE.【Conclusion】PpSS and PpSE genes were the new SS and SE family member in Pinus pumila.The acquisition of the genes will lay a foundation for further research on the regulatory mechanism in the sterol metabolism.

关 键 词：偃松 SS SE 克隆 

分 类 号：S791.24[农业科学—林木遗传育种]