Construction of yeast two-hybrid cDNA library of white-backed planthopper (Sogatella furcifera)

doi:10.3969/j.issn.1004-1524.2017.12.14

›› 2017, Vol. 29 ›› Issue (12): 2060-2067.DOI: 10.3969/j.issn.1004-1524.2017.12.14

• Plant Protection • Previous Articles Next Articles

Construction of yeast two-hybrid cDNA library of white-backed planthopper (Sogatella furcifera)

XUE Jin^{1, 2}, LI Jing², YANG Jian², TANG Yan^{1, 3}, LIANG Yao¹, CHEN Jianping^{2, *}, ZHANG Hengmu^{2, *}

1. Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Institute of Virology, College of Plant Protection, Hunan Agricultural University, Changsha 410128, China;
2. Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
3.College of Orient Science & Technology, Hunan Agricultural University, Changsha 410128, China

Received:2017-05-09 Online:2017-12-20 Published:2018-01-08

Abstract

Abstract: White-backed planthopper (Sogatella furcifera Horváth) is a major migratory pest that feed on rice plants and transmit Southern rice black-streaked dwarf virus (SRBSDV) in a persistent manner. In order to screen interactions between the genes of WBPH and SRBSDV, a cDNA expression library of WBPH was firstly constructed into the yeast plasmids using Gateway technique in this study. The results showed that the primary library consisted of 1.72×10⁷ CFU (colony-forming units), the sizes of most inserts were 500-1 500 bp in length, and the recombinant frequency was about 95.8%; the titer of secondary cDNA library was 1.73×10⁷ CFU, the sizes of most inserts were 400-2 000 bp in length, and the recombinant frequency was 100%; the titer assays showed that the cDNA library was approximately 5.09×10⁷ CFU for yeast two-hybrid cDNA library of Y187; PCR assays showed that the average length of insertions were 750-2 000 bp and the recombinant frequency of the cDNA library was high up to 100%, random sequence analysis of 36 clones showed that 22 of the clones were different in GenBank. These results consistently indicated that the cDNA library could be used for screening interactions in the yeast two hybrid experiments, which laid the foundation for studying on the interactions between white-backed planthopper and SRBSDV.

Key words: white-backed planthopper (Sogatella furcifera Horváth), Southern rice black-streaked dwarf virus (SRBSDV), Gateway technique, yeast two-hybrid, cDNA library

CLC Number:

S435.11

XUE Jin, LI Jing, YANG Jian, TANG Yan, LIANG Yao, CHEN Jianping, ZHANG Hengmu. Construction of yeast two-hybrid cDNA library of white-backed planthopper (Sogatella furcifera)[J]. , 2017, 29(12): 2060-2067.

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Construction of yeast two-hybrid cDNA library of white-backed planthopper (Sogatella furcifera)

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