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A protocol for protein extraction from recalcitrant tissues of grapevine (Vitis vinifera L.) for proteome analysis

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Title A protocol for protein extraction from recalcitrant tissues of grapevine (Vitis vinifera L.) for proteome analysis
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Creator Satisha Jogaiah, Anuradha Upadhyay, Smita R. Maske and Manisha P. Shinde
 
Subject Grapes, phenol extraction, protein extraction, proteomics, 2D electrophoresis
 
Description Not Available
The difficulty in obtaining high quality proteins from recalcitrant plant species like grape (Vitis vinifera L.) is mainly due
to low concentration of proteins, high activity of proteases and high level of interfering compounds, such as, pigments,
polyphenols, tannins, flavonoids etc. Utilization of 2-dimentional electrophoresis (2DE) technique in proteome analysis
largely depends on more efficient and optimized preparation of samples, which is regarded as one of the most important step
for obtaining reliable information on proteins. In the present study, four different previously published protein extraction
protocols and commercial protein extraction kits were compared to identify the most suitable protocol for protein extraction
from different grape tissues, such as, leaves, buds, clusters and berries. Of the five protocols compared, resolution of protein
in 1st dimension and 2nd dimension gel was improved with modified trichloro acetic acid/acetone precipitation with phenol
extraction method. This method also yielded higher protein concentration, greater spot resolution with minimal streaking on
2DE gels of grape leaf protein. The same protocol also yielded higher quality protein from other grape tissues, such as, buds,
berries and roots. Thus modified tricholoro acetic acid/acetone precipitation with phenol extraction method will be suitable
for proteome analysis of different grapevine tissues.
Not Available
 
Date 2019-03-21T07:13:18Z
2019-03-21T07:13:18Z
2015-10-01
 
Type Research Paper
 
Identifier Not Available
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/17603
 
Relation Not Available;
 
Publisher Indian Journal of Biotechnology