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Isolation and characterization of ge...

Manocha, Marcus M. S.

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Isolation and characterization of genomic DNA sequences that enhance the stability of plasmid DNA in mammalian cells.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Isolation and characterization of genomic DNA sequences that enhance the stability of plasmid DNA in mammalian cells./
Author:	Manocha, Marcus M. S.
Description:	144 p.
Notes:	Source: Masters Abstracts International, Volume: 45-01, page: 0224.
Contained By:	Masters Abstracts International45-01.
Subject:	Biology, Genetics. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=MR18001
ISBN:	9780494180013

Isolation and characterization of genomic DNA sequences that enhance the stability of plasmid DNA in mammalian cells.
Manocha, Marcus M. S.

Isolation and characterization of genomic DNA sequences that enhance the stability of plasmid DNA in mammalian cells. - 144 p.

Source: Masters Abstracts International, Volume: 45-01, page: 0224.

Thesis (M.Sc.)--Brock University (Canada), 2006.

The ease of production and manipulation has made plasmid DNA a prime target for its use in gene transfer technologies such as gene therapy and DNA vaccines. The major drawback of plasmid however is its stability within mammalian cells. Plasmid DNA is usually lost by cellular mechanisms or as a result of mitosis by simple dilution. This study set out to search for mammalian genomic DNA sequences that would enhance the stability of plasmid DNA in mammalian cells.

ISBN: 9780494180013Subjects--Topical Terms:

1017730
Biology, Genetics.

Isolation and characterization of genomic DNA sequences that enhance the stability of plasmid DNA in mammalian cells.
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020 $a 9780494180013
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040 $a UMI $c UMI
100 1 $a Manocha, Marcus M. S. $3 1922921
245 1 0 $a Isolation and characterization of genomic DNA sequences that enhance the stability of plasmid DNA in mammalian cells.
300 $a 144 p.
500 $a Source: Masters Abstracts International, Volume: 45-01, page: 0224.
502 $a Thesis (M.Sc.)--Brock University (Canada), 2006.
520 $a The ease of production and manipulation has made plasmid DNA a prime target for its use in gene transfer technologies such as gene therapy and DNA vaccines. The major drawback of plasmid however is its stability within mammalian cells. Plasmid DNA is usually lost by cellular mechanisms or as a result of mitosis by simple dilution. This study set out to search for mammalian genomic DNA sequences that would enhance the stability of plasmid DNA in mammalian cells.
520 $a Creating a plasmid based genomic DNA library, we were able to screen the human genome by transfecting the library into Human Embryonic Kidney (HEK 293) Cells. Cells that contained plasmid DNA were selected, using G418 for 14 days. The resulting population was then screened for the presence of biologically active plasmid DNA using the process of transformation as a detector.
520 $a A commercially available plasmid DNA isolation kit was modified to extract plasmid DNA from mammalian cells. The standardized protocol had a detection limit of ~0.6 plasmids per cell in one million cells. This allowed for the detection of 45 plasmids that were maintained for 32 days in the HEK 293 cells. Sequencing of selected inserts revealed a significantly higher thymine content in comparison to the human genome. Sequences with high A/T content have been associated with Scaffold/Matrix Attachment Region (S/MAR) sequences in mammalian cells. Therefore, association with the nuclear matrix might be required for the stability of plasmids in mammalian cells.
590 $a School code: 1476.
650 4 $a Biology, Genetics. $3 1017730
650 4 $a Biology, Cell. $3 1017686
690 $a 0369
690 $a 0379
710 2 0 $a Brock University (Canada). $3 1018667
773 0 $t Masters Abstracts International $g 45-01.
790 $a 1476
791 $a M.Sc.
792 $a 2006
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=MR18001