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

doi:10.7668/hbnxb.2015.06.010

Abstract

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

CLC Number:

S432.4

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.

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