This HTML5 document contains 21 embedded RDF statements represented using HTML+Microdata notation.

The embedded RDF content will be recognized by any processor of HTML5 Microdata.

Subject Item

n2:w

rdf:type

rdac:C10001 bibo:Article

dcterms:isPartOf

n10:w

dcterms:subject

n14:19 n14:methodsonline

dcterms:title

Minimal cross-recombination between wild-type and loxP511 sites in vivo facilitates truncating both ends of large DNA inserts in pBACe3.6 and related vectors

rdaw:P10072

n7:web n7:print

vivo:relatedBy

n4:1 n4:4 n4:2 n4:3 n4:5

marcrel:aut

n2:srivastavadeepakk n2:shakesleighcrafta n2:harewoodkenr n2:garlanddouglasm n15:7ac75f3947c9fa678d19b901dd3abfd5

dcterms:abstract

Contrary to several earlier reports, we find that cross-recombination between wild-type and the mutant loxP511 sites is <0.5% of that between two wild-type sites if Cre protein is expressed by phage P1 during an infection. The finding enabled us to develop a procedure to truncate DNA progressively from both ends of large genomic inserts flanked by these two loxP sites in pBACe3.6 and related vectors with transposons carrying either a wild-type or a loxP511 sequence. Newly constructed loxP511 transposons contained either a kanamycin resistance gene or no marker. Insert DNA ends in deletions were sequenced with primers unique to each transposon-end remaining after the respective recombination. End-sequencing 223 deletions confirmed that the low level of cross-recombination, observed between those sites during the P1 transductions, does not complicate the procedure: truncations from the unintended end of genomic inserts did not occur. Multiple BACs pooled together could also be processed in a single tube to make end-deletions. This deletion technology, utilizing the very minimal cross-recombination between the mutant and wild-type loxP sites of most BAC clones in the public domain and a heterologous one inserted as a transposon, should facilitate functionally mapping long-range gene regulatory sequences and help to isolate genes with defined functional boundaries in numerous projects including those of therapeutic interest.

hub:articleType

n12:other

hub:isPartOfThisJournal

n13:w