Rapid and efficient genetic transformation of the green microalga Chlorella vulgaris

Rapid and efficient genetic transformation of the green microalga Chlorella vulgaris Chlorella is a green unicellular alga that has a wide range of future biotechnical applications such as production of pharmaceuticals and biodiesel. Efficient genetic transformation of Chlorella vulgaris has been difficult due to technical limitations. In this study, an efficient and reliable transformation system of electroporation was established using two different reporter genes. First, C. vulgaris cell wall was digested with enzyme mixture for preparing protoplasts. The optimal transformation efficiency was 1.67 × 104 ± 0.083 cfu μg−1 plasmid under the following conditions: 2 × 106 cells mL−1 of growing culture; 655 V pulse voltage with 3.4 ms pulse width. After transformation, green and cyan fluorescence were observed from transgenic C. vulgaris harboring gfp (green fluorescent protein) gene of pCAMBIA1302 and cfp (cyan fluorescent protein) gene of pSK397, respectively, using laser confocal microscope. RT-PCR analysis as well as Southern blot confirmed the integration of reporter gene at the molecular level. This efficient transformation system of C. vulgaris would be valuable for the production of recombinant proteins in the future. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Applied Phycology Springer Journals

Rapid and efficient genetic transformation of the green microalga Chlorella vulgaris

Loading next page...
 
/lp/springer_journal/rapid-and-efficient-genetic-transformation-of-the-green-microalga-T06RqbQIT7
Publisher
Springer Journals
Copyright
Copyright © 2018 by Springer Science+Business Media B.V., part of Springer Nature
Subject
Life Sciences; Plant Sciences; Freshwater & Marine Ecology; Plant Physiology; Ecology
ISSN
0921-8971
eISSN
1573-5176
D.O.I.
10.1007/s10811-018-1396-3
Publisher site
See Article on Publisher Site

Abstract

Chlorella is a green unicellular alga that has a wide range of future biotechnical applications such as production of pharmaceuticals and biodiesel. Efficient genetic transformation of Chlorella vulgaris has been difficult due to technical limitations. In this study, an efficient and reliable transformation system of electroporation was established using two different reporter genes. First, C. vulgaris cell wall was digested with enzyme mixture for preparing protoplasts. The optimal transformation efficiency was 1.67 × 104 ± 0.083 cfu μg−1 plasmid under the following conditions: 2 × 106 cells mL−1 of growing culture; 655 V pulse voltage with 3.4 ms pulse width. After transformation, green and cyan fluorescence were observed from transgenic C. vulgaris harboring gfp (green fluorescent protein) gene of pCAMBIA1302 and cfp (cyan fluorescent protein) gene of pSK397, respectively, using laser confocal microscope. RT-PCR analysis as well as Southern blot confirmed the integration of reporter gene at the molecular level. This efficient transformation system of C. vulgaris would be valuable for the production of recombinant proteins in the future.

Journal

Journal of Applied PhycologySpringer Journals

Published: Jan 16, 2018

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 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

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