Expression, Purification and Binding Activity of Ripening Inhibitor (RIN) protein of Tomato Fruit

DOI

10.2991/icaees-15.2015.244How to use a DOI?

Keywords

Tomato; RIN protein; Prokaryotic expression; Protein purification; EMSA

Abstract

RIN protein plays a very important role in tomato fruit growth, development and ripening process. In order to study the regulation mechanism of RIN transcription factor in tomato fruit, the high purity RIN protein was firstly needed to get. In the study, RIN protein was efficiently expressed through the pET-RIN prokaryotic expression vector, 1.0 mmol/L IPTG induction 3 h at 37ºC. The recombinant protein RIN was mainly in inclusion body form 12% SDS-PAGE analysis. High purity of HIS-RIN fusion protein was gotten by affinity chromatography purification with the Ni Sepharose 6 Fast Flow. After renaturation of RIN protein, the binding activity of RIN protein was identified. The results showed that the renaturation of RIN protein was successfully obtained, and RIN transcription factor has the ability to combine the promoter of LeEXP1 and TBG4 gene. So the protein can be used for further molecular biology research.

Copyright

© 2015, the Authors. Published by Atlantis Press.

Open Access

This is an open access article distributed under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).

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TY  - CONF
AU  - Ling Li
AU  - Lu Liang
AU  - Yunxi Liu
AU  - Yanxiu Liu
AU  - Xiaodan Li
AU  - Tieling Liu
PY  - 2015/07
DA  - 2015/07
TI  - Expression, Purification and Binding Activity of Ripening Inhibitor (RIN) protein of Tomato Fruit
BT  - Proceedings of the 3rd International Conference on Advances in Energy and Environmental Science 2015
PB  - Atlantis Press
SP  - 1309
EP  - 1312
SN  - 2352-5401
UR  - https://doi.org/10.2991/icaees-15.2015.244
DO  - 10.2991/icaees-15.2015.244
ID  - Li2015/07
ER  -