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Molecular Cloning and Analysis of Expression Levels of Buchnera groEL of Aphis glycines(Hemiptera: Aphididae)(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2015年06期
Page:
964-971
Research Field:
Publishing date:

Info

Title:
Molecular Cloning and Analysis of Expression Levels of Buchnera groEL of Aphis glycines(Hemiptera: Aphididae)
Author(s):
XU Zhong-xin;  WANG Hong;  ZHAO Kui-jun;  HAN Lan-lan
College of Agriculture, Northeast Agricultural University, Harbin 150030, China
Keywords:
Aphis glycines; soybean mosaic virus; Buchnera; GroEL; Real-time quantitative PCR
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2015.06.0964
Abstract:
In order to probe into the mechanism underlying interaction between propagation of mosaic virus (soybean mosaic virus, SMV) by ?Aphis glycines and its Buchnera-producing GroEL protein. The full-length groEL sequence was amplified from the Buchnera of Aphis glycines by RT-PCR technique, and the gene was recombined into pET21b and expressed in prokaryotic expression system.Meanwhile purified recombinant protein was obtained by Ni-Agarose His affinity chromatography, the content of SMV in Aphis glycinesat different time and the changes of expression levels of groEL gene after feeding SMV were analyzed using Real-time quantitative PCR technology.The sequence analysis showed that the full-length Buchnera groEL sequence of Aphis glycines is 1 647 bp in length encoding 548 amino acid residues with the predicted molecular weight 69 kDa and pI 5.24.The built recombinant vector pET21b-GroEL was expressed in prokaryotic expression system，and the recombinant protein was determinded as the target protein using western-blot analysis. The Buchnera groEL gene expression levels of Aphis glycines was increased significantly with the extension of time of feeding SMV, the expression level of Buchnera groEL gene expression content in wingless and winged adults were significantly higher than the other four nymphs periods. All these results above showed that Buchneragro EL gene of Aphis glycines was expressed stably and correctly in E.coli, and the expression of Buchneragro EL was induced in Aphis glycines after feeding on SMV, which increases the possibility of SMV spread in soybean field.

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Last Update: 2016-01-05