感染重组型大豆花叶病毒本氏烟的酵母双杂文库构建及评价

doi:10.7668/hbnxb.2015.06.010

华北农学报 ›› 2015, Vol. 30 ›› Issue (6): 66-69. doi: 10.7668/hbnxb.2015.06.010

所属专题： 油料作物；

感染重组型大豆花叶病毒本氏烟的酵母双杂文库构建及评价

林静^1,2, 杨永庆¹, 侯文焕¹, 史晓蕾¹, 张孟臣¹, 谢令琴², 杨春燕¹

1. 河北省农林科学院粮油作物研究所, 国家大豆改良中心石家庄分中心, 农业部黄淮海大豆生物学与遗传育种重点实验室, 河北省作物遗传育种实验室, 河北石家庄 050035;
2. 河北农业大学农学院, 河北保定 071001

收稿日期:2015-10-08 出版日期:2015-12-28
作者简介:林静(1991-),女,福建龙岩人,在读硕士,主要从事大豆抗病育种研究.
基金资助:
国家自然科学基金项目(31401409;31201234;31471522);国家转基因生物新品种培育重大专项(2014ZX0800402B;2014ZX08004001); 河北省科技支撑计划项目(14226309D)

Construction and Evaluation of Y2H Library forNicotiana benthamiana Infection with Recombined Soybean Mosaic Virus

LIN Jing^1,2, YANG Yong-qing¹, HOU Wen-huan¹, SHI Xiao-lei¹, ZHANG Meng-chen¹, XIE Ling-qin², YANG Chun-yan¹

1. Institute of Cereal and Oil Crops, Hebei Academy of Agriculture and Forestry Sciences, National Soybean Improvement Center Shijiazhuang Sub-center, Huanghuaihai Key Laboratory of Biology and Genetic Improvement of Soybean, Ministry of Agriculture, Hebei Laboratory of Crop Genetics and Breeding, Shijiazhuang 050035, China;
2. College of Agronomy, Agricultural University of Hebei, Baoding 071001, China

Received:2015-10-08 Published:2015-12-28

摘要/Abstract

摘要： 为了建立以本氏烟为模式作物研究SMV侵染机制的平台,以可系统侵染本氏烟的重组型SMV分离物HB-RS为毒源,构建感染HB-RS本氏烟的酵母双杂cDNA文库.利用SMART技术获得本氏烟的双链cDNA文库,通过双酶切手段将cDNA文库构建至pGADT7-SfiI载体上.检测结果显示,文库容量约为3.2×10⁶ cfu,插入cDNA片段长度为0.75~2.00 kb,平均长度大于1.00 kb,且多态性丰富,表明文库质量较好.该酵母双杂文库的构建为钓取与SMV互作的寄主因子和系统性研究SMV侵染机制奠定了物质基础.

关键词: 重组型大豆花叶病毒, 本氏烟, 酵母双杂交文库

Abstract: In order to establish a platform for studying on the mechanism of SMV infection on the model crop of Nicotiana benthamiana.In this study,a recombinant SMV isolate HB-RS,which could systematically infect Nicotiana benthamiana ,was used as virus source to construct a yeast two hybrid cDNA library of Nicotiana benthamiana with HB-RS infection.Double stranded cDNA library of the Nicotiana benthamiana were obtained through SMART technology and cDNA library were cloned into pGADT7-SfiI vector by double enzyme digestion method.Testing result showed that the cDNA library contained more than 3.2×10⁶cfu and the cDNA fragments length inserted were from 0.75-2.00 kb.The fragment was polymorphism and average length was more than 1.00 kb indicating that the cDNA library was high quality.In all,cDNA library of the yeast two hybrids provided a material basis for screening host factor with SMV proteins and systematic studying of SMV infection mechanism.

Key words: Recombinant soybean mosaic virus, Nicotiana benthamiana, Yeast two hybrid cDNA library

中图分类号:

S432.4

林静, 杨永庆, 侯文焕, 史晓蕾, 张孟臣, 谢令琴, 杨春燕. 感染重组型大豆花叶病毒本氏烟的酵母双杂文库构建及评价[J]. 华北农学报, 2015, 30(6): 66-69. doi: 10.7668/hbnxb.2015.06.010.

LIN Jing, YANG Yong-qing, HOU Wen-huan, SHI Xiao-lei, ZHANG Meng-chen, XIE Ling-qin, YANG Chun-yan. Construction and Evaluation of Y2H Library forNicotiana benthamiana Infection with Recombined Soybean Mosaic Virus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(6): 66-69. doi: 10.7668/hbnxb.2015.06.010.

http://www.hbnxb.net/CN/Y2015/V30/I6/66

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