A rapid method for targeted modification and screening of recombinant bacterial artificial chromosome.
Modification of bacterial artificial chromosomes (BACs) has been a useful method to produce genomic DNA fragments for studying gene expression and function in vitro and in vivo. The original technique involved restrictions for BAC modification and required multiple cloning steps to target sequences into the shuttle vector. Selection and screening of BAC recombinants was accomplished by drug resistance and Southern blotting. We have developed a PCR-based method for producing the modified shuttle vectors and for screening for BACs carrying homologous integrants. The combination of these techniques allows for rapid and easy targeted BAC sequence deletion or insertion.
Misulovin
Z
Z
Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10021-6399, USA.
Yang
X W
XW
Yu
W
W
Heintz
N
N
Meffre
E
E
D001483:Base Sequence; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D018390:Gene Targeting; D005821:Genetic Techniques; D005822:Genetic Vectors; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D017384:Sequence Deletion
D017931:DNA Primers