scholarly journals A Low-Copy-Number Plasmid for Retrieval of Toxic Genes from BACs and Generation of Conditional Targeting Constructs

2012 ◽  
Vol 54 (2) ◽  
pp. 504-514 ◽  
Author(s):  
Giyoun Na ◽  
Andrew Wolfe ◽  
CheMyong Ko ◽  
Hyesook Youn ◽  
Young-Min Lee ◽  
...  
2004 ◽  
Vol 70 (4) ◽  
pp. 2508-2513 ◽  
Author(s):  
Satoshi Tomita ◽  
Kenji Tsuge ◽  
Yo Kikuchi ◽  
Mitsuhiro Itaya

ABSTRACT A method for positional cloning of the Bacillus subtilis genome was developed. The method requires a set of two small DNA fragments that flank the region to be copied. A 38-kb segment that carries genes ppsABCDE encoding five enzymes for antibiotic plipastatin synthesis and another genome locus as large as 100 kb including one essential gene were examined for positional cloning. The positional cloning vector for ppsABCDE was constructed using a B. subtilis low-copy-number plasmid that faithfully copied the precise length of the 38-kb DNA in vivo via the recombinational transfer system of this bacterium. Structure of the copied DNA was confirmed by restriction enzyme analyses. Furthermore, the unaltered structure of the 38-kb DNA was demonstrated by complementation of a ppsABCDE deletion mutant.


1988 ◽  
Vol 51 (1) ◽  
pp. 71-74 ◽  
Author(s):  
Kevin Kendall ◽  
John Cullum

SummaryWe constructed a cloning vector (pMT603) based on the low copy number plasmid SCP2*. pMT6O3 is unstable because it lacks the SCP2* stability region and carries the selectable marker thiostrepton-resistance and a tyrosinase gene which results in melanin production. This allows easy testing of plasmid stability and we demonstrated its usefulness by cloning a plasmid stability function.


1982 ◽  
Vol 150 (3) ◽  
pp. 1234-1243 ◽  
Author(s):  
W Firshein ◽  
P Strumph ◽  
P Benjamin ◽  
K Burnstein ◽  
J Kornacki

Gene ◽  
1984 ◽  
Vol 28 (1) ◽  
pp. 45-54 ◽  
Author(s):  
Jens Erik Løve Larsen ◽  
Kenn Gerdes ◽  
Janice Light ◽  
Søren Molin

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