DESIGNING AND CONSTRUCTING A PLASMID WITH MITOCHONDRIAL ORIGIN OF REPLICATION AND EVALUATING ITS REPLICATION IN HUMAN CELL LINES

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

AMINZADEH GOUHARI S.; KHANAHMAD H.; KARIMIPOUR MORTEZA; YAGHMAEI PARICHEHR; KARIMI ZARE S.; JAMSHIDI FATEMEH

Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: Internal Medicine Today Year:2009 | Volume:15 | Issue:3 Page(s): 45-56

Download Full-Text

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Persian Verion

View:

1,112

Download:

Cites:

Information Journal Paper

Title

DESIGNING AND CONSTRUCTING A PLASMID WITH MITOCHONDRIAL ORIGIN OF REPLICATION AND EVALUATING ITS REPLICATION IN HUMAN CELL LINES

Author(s)

Keywords

GENE THERAPYQ2

NON-VIRAL VECTORQ3

MITOCHONDRIAL ORIGIN OF REPLICATIONQ3

Abstract

Background and Aim: In order to achieve a safe vector with ability to replicate autonomously in human cells by human transfactors, a recombinant plasmid with human mitochondrial origins of replication was constructed. In contrast to lentiviral and adenoviral vectors, this plasmid does not integrate into the host genome and replicates stably. Materials and Methods: Both human mitochondrial origins of replication and gfp fragments were amplified by PCR, cloned into pTZ57T/A. Hygromycin resistance gene was digested from pFBGGT. Then, four DNA fragments were subcloned into pBGGT plasmid. All steps of cloning were checked by PCR, restricted analysis and sequencing. HEK293 and CHO cell lines were transfected by final plasmid (pEU). Transfected cells were checked by Fluorescence Microscope daily during 40 days. pEU and genomic DNA were extracted from transfected HEK293 treated with hygromycin. Five overlap PCRs were performed on these products to check presence of circular plasmid in transfected HEK293.Results: All steps of cloning were confirmed. Data showed that pEU did not replicate in transfected cells. Conclusion: As recombinant plasmids with both mitochondrial origins of replication did not replicate in transfected cell lines, it seems that we need to provide similar conditions of mitochondrial DNA replication in order to replicate the above vector in future experiments.

Cites

No record.

References

No record.

Cite

APA: Copy

KHANAHMAD, H., KARIMIPOUR, MORTEZA, YAGHMAEI, PARICHEHR, KARIMI ZARE, S., JAMSHIDI, FATEMEH, & AMINZADEH GOUHARI, S.. (2009). DESIGNING AND CONSTRUCTING A PLASMID WITH MITOCHONDRIAL ORIGIN OF REPLICATION AND EVALUATING ITS REPLICATION IN HUMAN CELL LINES. OFOGH-E-DANESH, 15(3), 45-56. SID. https://sid.ir/paper/68796/en

Vancouver: Copy

KHANAHMAD H., KARIMIPOUR MORTEZA, YAGHMAEI PARICHEHR, KARIMI ZARE S., JAMSHIDI FATEMEH, AMINZADEH GOUHARI S.. DESIGNING AND CONSTRUCTING A PLASMID WITH MITOCHONDRIAL ORIGIN OF REPLICATION AND EVALUATING ITS REPLICATION IN HUMAN CELL LINES. OFOGH-E-DANESH[Internet]. 2009;15(3):45-56. Available from: https://sid.ir/paper/68796/en

IEEE: Copy

H. KHANAHMAD, MORTEZA KARIMIPOUR, PARICHEHR YAGHMAEI, S. KARIMI ZARE, FATEMEH JAMSHIDI, and S. AMINZADEH GOUHARI, “DESIGNING AND CONSTRUCTING A PLASMID WITH MITOCHONDRIAL ORIGIN OF REPLICATION AND EVALUATING ITS REPLICATION IN HUMAN CELL LINES,” OFOGH-E-DANESH, vol. 15, no. 3, pp. 45–56, 2009, [Online]. Available: https://sid.ir/paper/68796/en

Related Journal Papers