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Caption a, Targeting strategy to generate the Col14a1tm1Debi allele. Functional domains of type XIV collagen and genomic structure of mouse Col14a1 are shown in the upper part of the diagram. In the gene-targeting vector, exon 4 was split into two parts, and a stop codon was inserted into the 5'-half of E4 by PCR. Genomic sequences of exon 3 through the first part of exon 4 and the second part of exon 4 through exon 5 were chosen as 5'- and 3'-targeting arms. Both arms are about 3 kb in size. Neomycin-resistant gene was inserted between the two arms, reversely (black arrows show transcription direction). Probes (short color rectangles) and restriction enzymes used for genomic DNA digestion and Southern blotting are indicated in the diagram. Green arrows represent gene-specific primers for genotyping. Primer 1 (P1) and Primer 2 (P2), from the second part of exon 4 and intron 4, respectively, produce a PCR product from the wild-type and mutant alleles. Primer 3 (P3) from the Neo gene, produces a DNA fragment only from the mutant allele when used with P2. The translation termination codon, which is located in FNIII-1, is marked on the protein structure diagram. The anti-type XIV collagen antibody epitope also is marked in this diagram.
Copyright This image is from Ansorge HL, J Biol Chem 2009 Mar 27;284(13):8427-38 and is displayed with the permission of the American Society for Biochemistry and Molecular Biology who owns the Copyright. Full text from JBC. J:148619
Associated
Alleles
Symbol Name
Col14a1tm1Debi collagen, type XIV, alpha 1; targeted mutation 1, David E Birk

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last database update
12/10/2024
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