The Construction of Mutant <i>eIF4E</i> in Papaya

ZHANG Dong-dong; YAN Pu; SHEN Wen-tao; LI Xiao-ying; ZHOU Peng

doi:10.15886/j.cnki.rdswxb.2012.02.014

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Mutations in or loss of the eukaryotic translation initiation factor 4E（eIF4E） can affect infections of viruses to plants,which leads to resistance of the plants against the viruses.The interaction between PRSV-Vpg and Cp-eIF4E（eIF4E in Papaya） has been verified.In this study,six point mutations were determined by protein sequence alignment and analysis of homology modeling,and the point mutations were produced in Cp-eIF4E by overlap extension PCR.Then,the mutant Cp-eIF4E genes were connected into two sets of expressive vectors,yeast two-hybrid system and bimolecular fluorescence complementation,which lays a foundation for the follow-up verification of interaction and resistance in the plants.

The Construction of Mutant eIF4E in Papaya

1. College of Agronomy, Hainan University, Haikou 570228, China;

2. Institute of Tropical Bioscience and Biotechnology/Analysis & Testing Center, Chinese Acaderny of Tropical Agricultural Sciences, Haikou 571101, China

Received Date: 2012-02-14

Key words:

eIF4E /
point mutation /
Vpg /
interaction

Abstract: Mutations in or loss of the eukaryotic translation initiation factor 4E（eIF4E） can affect infections of viruses to plants,which leads to resistance of the plants against the viruses.The interaction between PRSV-Vpg and Cp-eIF4E（eIF4E in Papaya） has been verified.In this study,six point mutations were determined by protein sequence alignment and analysis of homology modeling,and the point mutations were produced in Cp-eIF4E by overlap extension PCR.Then,the mutant Cp-eIF4E genes were connected into two sets of expressive vectors,yeast two-hybrid system and bimolecular fluorescence complementation,which lays a foundation for the follow-up verification of interaction and resistance in the plants.

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[1]	WITTMANN S,CHATEL H,FORTIN M G,et al.Interaction of the viral protein genome linked of Turnip mosaic potyvirus with the translational Eukaryotic initiation factor(iso)4E of Arabidopsis thaliana using the yeast two-hybrid system[J].Virolo-gy,1997,234:84-92.
[2]	RUFFEL S,DUSSAULT M H,PALLOIX A,et al.A natural recessive resistance gene against Potato virus Y in peppercorre-sponds to the eukaryotic initiation factor4E(eIF4E)[J].The Plant Journal,2002,32(6):1067-1075.
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[7]	吴金燕,沈文涛,言普,等.番木瓜eIF4E和eIFiso4E克隆及其与PRSV-VPg互作研究[D].海口:海南大学,2010.
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The Construction of Mutant eIF4E in Papaya

doi: 10.15886/j.cnki.rdswxb.2012.02.014

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The Construction of Mutant eIF4E in Papaya

doi: 10.15886/j.cnki.rdswxb.2012.02.014

1. College of Agronomy, Hainan University, Haikou 570228, China;

2. Institute of Tropical Bioscience and Biotechnology/Analysis & Testing Center, Chinese Acaderny of Tropical Agricultural Sciences, Haikou 571101, China

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The Construction of Mutant eIF4E in Papaya

doi: 10.15886/j.cnki.rdswxb.2012.02.014

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Proportional views

Related

The Construction of Mutant eIF4E in Papaya

doi: 10.15886/j.cnki.rdswxb.2012.02.014

1. College of Agronomy, Hainan University, Haikou 570228, China; 2. Institute of Tropical Bioscience and Biotechnology/Analysis & Testing Center, Chinese Acaderny of Tropical Agricultural Sciences, Haikou 571101, China

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1. College of Agronomy, Hainan University, Haikou 570228, China;

2. Institute of Tropical Bioscience and Biotechnology/Analysis & Testing Center, Chinese Acaderny of Tropical Agricultural Sciences, Haikou 571101, China