Identification of VvEXP Gene from Grape Roots Infected by Phylloxera and Preparation of Polyclonal Antibody to Grape Expansin

doi:10.16420/j.issn.0513-353x.2015-0574

ACTA HORTICULTURAE SINICA ›› 2016, Vol. 43 ›› Issue (1): 15-24.doi: 10.16420/j.issn.0513-353x.2015-0574

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Identification of VvEXP Gene from Grape Roots Infected by Phylloxera and Preparation of Polyclonal Antibody to Grape Expansin

JI Xing-long1，HAN Ning2，ZHAI Heng1，and DU Yuan-peng1,*

1College of Horticulture Science and Engineering，Shandong Agricultural University，State Key Lab of Crop Biology，Tai’an，Shandong 271018，China；2College of Bioengineering，Qilu Technology of University，Ji’nan 250353，China

Online:2016-01-25 Published:2016-01-25

Abstract

Abstract:

Based on qRT-PCR method，an expansin（VvEXPA1）was acquired and investigated from infected Crimson Seedless grape and 1103P grape rootstock in present study. VvEXPA1 was cloned into prokaryotic expression vector pET32a after identification by enzymatic digestion and sequencing，and then the recombinant plasmid with VvEXPA1 was transformed into E. coli DE3. The results showed that the protein was highly expressed in E. coli and the molecular weight of the recombinant protein was 41 kD. The specific antibody was prepared by immunizing rabbit with the purified protein，and then detected by Western-blotting and ELISA. Finally，the high specific antibody was obtained and the titers of anti-VvEXPA1 antibody were 1︰51 200.

Key words: grape, phylloxera, expansin, VvEXPA1, poly-antibody preparation

CLC Number:

S 663.1

JI Xing-long1，HAN Ning2，ZHAI Heng1，and DU Yuan-peng1,*. Identification of VvEXP Gene from Grape Roots Infected by Phylloxera and Preparation of Polyclonal Antibody to Grape Expansin[J]. ACTA HORTICULTURAE SINICA, 2016, 43(1): 15-24.

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Identification of VvEXP Gene from Grape Roots Infected by Phylloxera and Preparation of Polyclonal Antibody to Grape Expansin

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