[1]王培,张家君,吕蒙蒙,等. 杉木ClLAR基因的克隆及遗传转化拟南芥[J].西北林学院学报,2019,34(5):1-9.[doi:10.3969/j.issn.1001-7461.2019.05.01]
 WANG Pei,ZHANG Jia-jun,LV Meng-meng,et al. Cloning of ClLAR Gene and Its Transformation in Arabidopsis thaliana[J].JOURNAL OF NORTHWEST FORESTRY UNIVERSITY,2019,34(5):1-9.[doi:10.3969/j.issn.1001-7461.2019.05.01]
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 杉木ClLAR基因的克隆及遗传转化拟南芥()
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《西北林学院学报》[ISSN:1001-7461/CN:61-1202/S]

卷:
第34卷
期数:
2019年第5期
页码:
1-9
栏目:
出版日期:
2019-09-30

文章信息/Info

Title:
 Cloning of ClLAR Gene and Its Transformation in Arabidopsis thaliana
文章编号:
1001-7461(2019)05-0001-09
作者:
 王培12张家君12吕蒙蒙12理挪12马志慧23陈宇12*林思祖12
 (1.福建农林大学 林学院,福建 福州 350002;2.国家林业局 杉木工程技术研究中心,福建 福州 350002;3.福建农林大学 生命科学学院,福建 福州 350002)
Author(s):
 WANG Pei12ZHANG Jia-jun12LV Meng-meng12LI Nuo12MA Zhi-hui23CHEN Yu12*LIN Si-zu12
 (1.College of Forestry,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China; 2.State Forestry Administration Engineering Research Center of Chinese Fir,Fuzhou 350002,Fujian,China; 3.College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China)
关键词:
 杉木ClLAR基因克隆序列分析载体构建遗传转化
Keywords:
 Chinese fir ClLAR gene cloning sequence analysis vector construction genetic transformation
分类号:
S791.27
DOI:
10.3969/j.issn.1001-7461.2019.05.01
文献标志码:
A
摘要:
 为探究杉木种子发育过程中无色花青素还原酶(LAR)基因在类黄酮生物合成途径中对原花青素(PA)合成的分子调控机制,以杉木种子为试验材料,通过RT-PCR结合RACE的方法成功克隆到杉木ClLAR基因全长,该基因的cDNA全长为1 625 bp,具有1个1 242 bp的开放阅读框(ORF),编码413个氨基酸。构建pCambia3301-ClLAR植物表达载体,运用冻融法将重组质粒转至农杆菌GV3101,通过花序侵染法获得拟南芥转基因植株,利用Basta溶液和PCR验证筛选出阳性苗,确定目的基因ClLAR已整合至拟南芥基因组中。研究结果为深入分析杉木ClLAR基因调控PA合成的分子机制和ClLAR蛋白功能奠定了基础。
Abstract:
 To explore the molecular regulation mechanism of leucocyanidin reductase (LAR) gene in the PA synthesis of the flavonoid biosynthesis pathway during the development of Chinese fir (Cunninghamia lanceolata) seeds,Chinese fir seeds were used as experimental materials,the ClLAR gene was cloned by RT-PCR combined with RACE.The results showed that the full length of ClLAR gene cDNA was 1 625 bp,it contained open reading frame (ORF) of 1 242 bp which encoded 413 amino acids.The plant expression vector of pCambia3301-ClLAR was constructed,and the recombinant plasmid was transferred to Agrobacterium tumefaciens GV3101 by freeze-thaw method.The transgenic plants of Arabidopsis thaliana were obtained by inflorescence infection method,and positive seedlings were screened by Basta solution and PCR.The results showed that the target gene had been integrated into the Arabidopsis genome.The results of this study laid the foundation for the further analysis of the molecular mechanism of ClLAR gene regulation of PA synthesis and the function of ClLAR protein in Chinese fir.

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备注/Memo

备注/Memo:
 收稿日期:2018-10-17修回日期:2019-05-22
基金项目:国家林业局杉木工程技术研究中心平台建设(ptjh130002);国家林业局杉木工程技术研究中心孵化基金(6213C011103)。
作者简介:王培,女,硕士,研究方向:林木遗传育种。E-mail:790646055@qq.com
*通信作者:陈宇,男,研究实习员,研究方向:林木遗传育种。E-mail:28811852@qq.com
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