Isolation and purification of expression product of Populus euphratica CPD (PeCPD) in Escherichia coli: A comparison of two different methods

Isolation and purification of expression product of Populus euphratica CPD (PeCPD) in Escherichia... An effective procedure for purifying the expression product of Populus euphratica constitutive photomorphogenesis and dwarf (PeCPD) gene in Escherichia coli had been developed. The product, which is a cytochrome P450 monooxygenase, participating in plant brassinosteroid (BR) biosynthesis was deduced. This method is based on the combination of differential centrifugation (DC) with preparative SDS-PAGE and below is named as DC-PSDS-PAGE. The target protein obtained by this method was apparently better in purity and yield than that obtained by Ni2+ chelate affinity chromatography (Ni2+-CAC). The target protein purity and yield was 95% and 37.897 mg/L of bacterial culture, respectively, in DC-PSDS-PAGE vs. 30% and 0.816 mg/L of bacterial culture in Ni2+-CAC. To the best of our knowledge, this procedure is reported for the first time. The reasons for selecting this method and its advantages are discussed in the paper. The purified protein can be used for raising antibody, which is necessary for in situ localization, and this will be a very important part of our subsequent experiments in studying the function of the target protein. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Plant Physiology Springer Journals

Isolation and purification of expression product of Populus euphratica CPD (PeCPD) in Escherichia coli: A comparison of two different methods

Loading next page...
 
/lp/springer_journal/isolation-and-purification-of-expression-product-of-populus-euphratica-Wqm9chGB1D
Publisher
SP MAIK Nauka/Interperiodica
Copyright
Copyright © 2012 by Pleiades Publishing, Ltd.
Subject
Life Sciences; Plant Physiology; Plant Sciences
ISSN
1021-4437
eISSN
1608-3407
D.O.I.
10.1134/S1021443712050068
Publisher site
See Article on Publisher Site

Abstract

An effective procedure for purifying the expression product of Populus euphratica constitutive photomorphogenesis and dwarf (PeCPD) gene in Escherichia coli had been developed. The product, which is a cytochrome P450 monooxygenase, participating in plant brassinosteroid (BR) biosynthesis was deduced. This method is based on the combination of differential centrifugation (DC) with preparative SDS-PAGE and below is named as DC-PSDS-PAGE. The target protein obtained by this method was apparently better in purity and yield than that obtained by Ni2+ chelate affinity chromatography (Ni2+-CAC). The target protein purity and yield was 95% and 37.897 mg/L of bacterial culture, respectively, in DC-PSDS-PAGE vs. 30% and 0.816 mg/L of bacterial culture in Ni2+-CAC. To the best of our knowledge, this procedure is reported for the first time. The reasons for selecting this method and its advantages are discussed in the paper. The purified protein can be used for raising antibody, which is necessary for in situ localization, and this will be a very important part of our subsequent experiments in studying the function of the target protein.

Journal

Russian Journal of Plant PhysiologySpringer Journals

Published: Aug 16, 2012

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 12 million articles from more than
10,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Unlimited reading

Read as many articles as you need. Full articles with original layout, charts and figures. Read online, from anywhere.

Stay up to date

Keep up with your field with Personalized Recommendations and Follow Journals to get automatic updates.

Organize your research

It’s easy to organize your research with our built-in tools.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve Freelancer

DeepDyve Pro

Price
FREE
$49/month

$360/year
Save searches from
Google Scholar,
PubMed
Create lists to
organize your research
Export lists, citations
Read DeepDyve articles
Abstract access only
Unlimited access to over
18 million full-text articles
Print
20 pages/month
PDF Discount
20% off