Construction of a plasmid vector containing epidermal growth factor receptor and C-Jun shRNA

Construction of a plasmid vector containing epidermal growth factor receptor and C-Jun shRNA The objective of this study was to construct a plasmid vector for EGFR-hm-1 and C-Junh-825 small interfering RNA (siRNA). EGFR-hm-1 and C-Jun-hm-825 oligonucleotide fragments were synthesized and short hairpin RNA (shRNA) were amplified by PCR. Plasmids were isolated from E. coli TOP10 bacterium by restriction enzyme digestion using pst1 and BamH1 and oligonucleotide fragments were cloned into the pSilencer plasmid containing the U6 promoter. Recombinant clones were generated by transforming JM109 competent cells with plasmid vectors containing shRNA molecules. 58 base-paired EGFR-hm-1 and 59 base-paired C-Jun-hm-825 oligonucleotide fragments were isolated. The fragments were 100% homologous with human sequences available on GenBank. The recombinant pSilencer1.0 vector containing a 58-bp EGFR-hm-1 and 59-bp C-Jun-hm-825 fragment was constructed. These vectors have the potential to be used as treatment to combat skin photoaging under UV exposure. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Dermatological Research Springer Journals

Construction of a plasmid vector containing epidermal growth factor receptor and C-Jun shRNA

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Publisher
Springer Berlin Heidelberg
Copyright
Copyright © 2018 by Springer-Verlag GmbH Germany, part of Springer Nature
Subject
Medicine & Public Health; Dermatology
ISSN
0340-3696
eISSN
1432-069X
D.O.I.
10.1007/s00403-017-1803-7
Publisher site
See Article on Publisher Site

Abstract

The objective of this study was to construct a plasmid vector for EGFR-hm-1 and C-Junh-825 small interfering RNA (siRNA). EGFR-hm-1 and C-Jun-hm-825 oligonucleotide fragments were synthesized and short hairpin RNA (shRNA) were amplified by PCR. Plasmids were isolated from E. coli TOP10 bacterium by restriction enzyme digestion using pst1 and BamH1 and oligonucleotide fragments were cloned into the pSilencer plasmid containing the U6 promoter. Recombinant clones were generated by transforming JM109 competent cells with plasmid vectors containing shRNA molecules. 58 base-paired EGFR-hm-1 and 59 base-paired C-Jun-hm-825 oligonucleotide fragments were isolated. The fragments were 100% homologous with human sequences available on GenBank. The recombinant pSilencer1.0 vector containing a 58-bp EGFR-hm-1 and 59-bp C-Jun-hm-825 fragment was constructed. These vectors have the potential to be used as treatment to combat skin photoaging under UV exposure.

Journal

Archives of Dermatological ResearchSpringer Journals

Published: Jan 20, 2018

References

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