Abstract:
:Escherichia coli-based artificial chromosomes have become important tools for physical mapping and sequencing in various genome projects. The lack of a general method to modify these large bacterial clones, however, has limited their utility in functional studies. We developed a simple method to modify bacterial artificial chromosomes directly in the recombination-deficient E. coli host strain by homologous recombination for in vivo studies. The IRES-LacZ marker gene was introduced into a 131 kb BAC containing the murine zinc finger gene, RU49. No rearrangements or deletions were detected in the modified BACs. Furthermore, transgenic mice were generated by pronuclear injection of the modified BAC, and germline transmission of the intact BAC has been obtained. Proper expression of the lacZ transgene in the brain has been observed, which could not be obtained with conventional transgenic constructs.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
Yang XW,Model P,Heintz Ndoi
10.1038/nbt0997-859subject
Has Abstractpub_date
1997-09-01 00:00:00pages
859-65issue
9eissn
1087-0156issn
1546-1696journal_volume
15pub_type
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