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A General Method for Direct DNA Vector Modification(PDF)

《南京师大学报(自然科学版)》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2007年04期
Page:
80-83
Research Field:
生命科学
Publishing date:

Info

Title:
A General Method for Direct DNA Vector Modification
Author(s):
Xie FengZhu LeiShang Guangdong
Jiangsu Key Laboratory for Biodiversity and Biotechnology,Nanjing 210046,China
Keywords:
recombineering three p ieces cloning conjugal transfer vector
PACS:
Q78
DOI:
-
Abstract:
Cloning one ormore DNA fragments into a vector is the one ofmost commonly used techniques in molecular biology. Classic cloning strategy, when dealt with comp licated cloning step s, like multip le fragments insertion, selective marker exchange, are often troubled by the choice of restriction enzymes, the base mutations caused by long fragment PCR, UV exposure and gel purification. We made use of the Red /ET technology to clone two fragments into the vector while remove the unnecessary vector part in a single step. This three p ieces cloningmanipulation is straightforward and convenient, with the potential to be a general cloning strategy for vector engineering.

References:

[ 1 ]  Zhang Y, Buchholz F, Muyrers J P, et al. A new logic for DNA engineering using recombination in Escherichia coli [ J ].Nat Genet, 1998, 20 (2) : 123-128.
[ 2 ]  Glaser S, Anastassiadis K, StewartA F. Current issues in mouse genome engineering[ J ]. Nat Genet, 2005, 37 (1) : 1 187-1193.
[ 3 ]  SarovM, Schneider S, PozniakovskiA. A recombineering p ipeline for functional genomics app lied to Caenorhabditis elegans[ J ]. NatMethods, 2006, 3 (10) : 839-844.
[ 4 ]  Wang J , SarovM, Rientjes J, et al. An imp roved recombineering app roach by adding RecA to lambda Red recombination[ J ]. Mol Biotechnol, 2006, 32 (1) : 43-53.
[ 5 ]  BiermanM, Logan R, O’Brien K, et al. Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Strep tomyces spp [ J ]. Gene, 1992, 116 (1) : 43-49.
[ 6 ]  Hosted T J , Baltz R H. Use of rp sL for dominance selection and gene rep lacement in trep tomyces roseosporus[ J ]. J Bacteriol, 1997, 179 (1) : 180-186.
[ 7 ]  Sambrook J, Fritsch E F, Maniatis T. Molecular Cloning:A LaboratoryManual[M ]. 2nd ed. New York: Cold Sp ring Harbor Laboratory Press, 1989.
[ 8 ]  Kieser T, BibbM J , ButtnerM J. Practical Strep tomyces Genetics[M ]. Norwich: The John Innes Foundation, 2000: 247-248.

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Last Update: 2013-05-05