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Volumn 272, Issue 5261, 1996, Pages 548-551

Altered growth and branching patterns in synpolydactyly caused by mutations in HOXD13

Author keywords

[No Author keywords available]

Indexed keywords

ALANINE;

EID: 0029871929     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.272.5261.548     Document Type: Article
Times cited : (498)

References (29)
  • 6
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    • W. J. Gehring et al , Cell 78, 211 (1994).
    • (1994) Cell , vol.78 , pp. 211
    • Gehring, W.J.1
  • 16
    • 15844367767 scopus 로고    scopus 로고
    • note
    • For sequencing of exon 1 of the HOXD13 gene the cosmid clone G2 (15), containing the homeobox genes HOXD10, HOXD11, HOXD12, HOXD13, and EVX2, was digested with Eco RI, and a 10-kb fragment containing the entire HOXD13 and the 5′ half of the HOXD12 gene was subcloned in the Eco RI site of pBluescript With this fragment as a template, the sense primer 5′-TTTGTATCAGGGQATGTGGC-3′ and the antisense primer 5′-CCACATCAGGAGA-CAGTATC-3′ were used to amplify exon 2 of the HOXD13 gene by PCR. A 1.3-kb Pst I fragment derived from the 10-kb Eco RI fragment hybridized with the PCR product and was therefore subcloned in the Pst I site of pBluescript and sequenced by dideoxynucleotide cycle sequencing. Because the 5′ end of the sequence of the Pst I fragment did not reach the translational start codon ATG of the HOXD13 gene, the primer 5′-ACATACGGCAGCTG-TAGTAG-3′, corresponding to a sequence 70 bp downstream of the 5′ Pst I site, was used for sequencing with the 10-kb Eco RI fragment as the template
  • 17
    • 15844368752 scopus 로고    scopus 로고
    • note
    • 2, 200 μM deoxynucleotide triphosphate (dNTP) without dGTP, 200 μM 7-deaza-2′-deoxyguanosine 5′-triphosphate, a final concentration of 0 5 μM for each primer, 10% dimethyl sulfoxide, and 0.2 U of Taq polymerase. The PCR products were analyzed on a 5% polyacrylamide gel under nondenaturing conditions. PCR products from affected individuals were subcloned into pCRII (invitrogen) and subjected to cycle sequencing
  • 22
    • 0027169185 scopus 로고
    • _, EMBO J 12, 2723 (1993).
    • (1993) EMBO J , vol.12 , pp. 2723
  • 23
    • 0027358721 scopus 로고
    • P. Dolle et a l, Cell 75, 431 (1993).
    • (1993) Cell , vol.75 , pp. 431
    • Dolle, P.1
  • 29
    • 15844391002 scopus 로고    scopus 로고
    • note
    • The cosmid clone G2, containing the HOXD13 gene, was provided by E Boncinelli We want to thank C Tabin for helpful discussions. Y. Pittel provided secretarial assistance Supported by NIH grants AR36819 and AR36820 (to B R.O.) and Deutsche Forschungsgemeinschaft (to S M.)


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