Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines
Caviar-producing fish with their economically valuable product are important in fisheries. The cDNA growth hormone (GH) of Beluga sturgeon (Huso huso) was constructed using total RNA from pituitary glands. To construct the recombinant and active lentiviruses carring GH gene, this DNA sequence was in...
Main Authors: | , , , , , |
---|---|
Format: | Article in Journal/Newspaper |
Language: | Persian |
Published: |
2014
|
Subjects: | |
Online Access: | http://aquaticcommons.org/24906/ http://aquaticcommons.org/24906/1/JMST_Volume%2013_Issue%201_Pages%201-10.pdf |
id |
ftaquaticcommons:oai:generic.eprints.org:24906 |
---|---|
record_format |
openpolar |
spelling |
ftaquaticcommons:oai:generic.eprints.org:24906 2023-05-15T15:41:47+02:00 Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines Mashjoor, S. Zolgharnain, H. Gardaneh, M. Salari Aliabadi, M.A. Qasemi, A. Azimi, Z. 2014 application/pdf http://aquaticcommons.org/24906/ http://aquaticcommons.org/24906/1/JMST_Volume%2013_Issue%201_Pages%201-10.pdf fa per http://aquaticcommons.org/24906/1/JMST_Volume%2013_Issue%201_Pages%201-10.pdf Mashjoor, S. and Zolgharnain, H. and Gardaneh, M. and Salari Aliabadi, M.A. and Qasemi, A. and Azimi, Z. (2014) Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines. Journal of Marine Science and Technology, 13(1), pp. 1-10. Biology Article PeerReviewed 2014 ftaquaticcommons 2020-02-27T09:32:02Z Caviar-producing fish with their economically valuable product are important in fisheries. The cDNA growth hormone (GH) of Beluga sturgeon (Huso huso) was constructed using total RNA from pituitary glands. To construct the recombinant and active lentiviruses carring GH gene, this DNA sequence was inserted into the cloning vector pTZ57R/T and subsequently cutted from pTZ57R/T by endonuclease enzyme and incorporated into lentivirus vector pNL-EGFP/CMV-WPRE on upstream of an IRES cassette. We also insert a reporter EGFP gene downstream of IRES so transfection and transduction steps can be traced. Using this vector plus virus packaging and envelope vectors, HEK293T cells was co-transfected by DNA-Lipofectamine complexes method. Cell supernatant full of virions was collected 48 hours later and concentrated using Amicon columns to obtain a high-titer virus stock. Nearly 1/5 of this stock was applied to a new batch of cultured HEK-293T. After 72h expression of EGFP gene was detected and the cells was collected for further analysis. Total RNA of these transduced cells was extracted and GH mRNA expression was revealed by RT-PCR. Results showed that, lentiviral vectors (LV) as a gene transfer system provide efficient delivery, integration and long-term expression by establishing a stable provirus in target cells and could be important tool in aquaculture and fisheries biotechnology research to increase the growth rate of farmed fish by transferring growth hormone (GH) transgenes into fish. Article in Journal/Newspaper Beluga Beluga* International Association of Aquatic and Marine Science Libraries and Information Centers (IAMSLIC): Aquatic Commons |
institution |
Open Polar |
collection |
International Association of Aquatic and Marine Science Libraries and Information Centers (IAMSLIC): Aquatic Commons |
op_collection_id |
ftaquaticcommons |
language |
Persian |
topic |
Biology |
spellingShingle |
Biology Mashjoor, S. Zolgharnain, H. Gardaneh, M. Salari Aliabadi, M.A. Qasemi, A. Azimi, Z. Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines |
topic_facet |
Biology |
description |
Caviar-producing fish with their economically valuable product are important in fisheries. The cDNA growth hormone (GH) of Beluga sturgeon (Huso huso) was constructed using total RNA from pituitary glands. To construct the recombinant and active lentiviruses carring GH gene, this DNA sequence was inserted into the cloning vector pTZ57R/T and subsequently cutted from pTZ57R/T by endonuclease enzyme and incorporated into lentivirus vector pNL-EGFP/CMV-WPRE on upstream of an IRES cassette. We also insert a reporter EGFP gene downstream of IRES so transfection and transduction steps can be traced. Using this vector plus virus packaging and envelope vectors, HEK293T cells was co-transfected by DNA-Lipofectamine complexes method. Cell supernatant full of virions was collected 48 hours later and concentrated using Amicon columns to obtain a high-titer virus stock. Nearly 1/5 of this stock was applied to a new batch of cultured HEK-293T. After 72h expression of EGFP gene was detected and the cells was collected for further analysis. Total RNA of these transduced cells was extracted and GH mRNA expression was revealed by RT-PCR. Results showed that, lentiviral vectors (LV) as a gene transfer system provide efficient delivery, integration and long-term expression by establishing a stable provirus in target cells and could be important tool in aquaculture and fisheries biotechnology research to increase the growth rate of farmed fish by transferring growth hormone (GH) transgenes into fish. |
format |
Article in Journal/Newspaper |
author |
Mashjoor, S. Zolgharnain, H. Gardaneh, M. Salari Aliabadi, M.A. Qasemi, A. Azimi, Z. |
author_facet |
Mashjoor, S. Zolgharnain, H. Gardaneh, M. Salari Aliabadi, M.A. Qasemi, A. Azimi, Z. |
author_sort |
Mashjoor, S. |
title |
Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines |
title_short |
Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines |
title_full |
Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines |
title_fullStr |
Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines |
title_full_unstemmed |
Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines |
title_sort |
cloning of the gh gene from the beluga sturgeon (huso huso) into a lentivial & none viral constructs and its expression in hek cell lines |
publishDate |
2014 |
url |
http://aquaticcommons.org/24906/ http://aquaticcommons.org/24906/1/JMST_Volume%2013_Issue%201_Pages%201-10.pdf |
genre |
Beluga Beluga* |
genre_facet |
Beluga Beluga* |
op_relation |
http://aquaticcommons.org/24906/1/JMST_Volume%2013_Issue%201_Pages%201-10.pdf Mashjoor, S. and Zolgharnain, H. and Gardaneh, M. and Salari Aliabadi, M.A. and Qasemi, A. and Azimi, Z. (2014) Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines. Journal of Marine Science and Technology, 13(1), pp. 1-10. |
_version_ |
1766374674644598784 |