Skip Navigation

This Article
Right arrow Print PDF (2941K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (117)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Verweij, C. L.
Right arrow Articles by Pannekoek, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Verweij, C. L.
Right arrow Articles by Pannekoek, H.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1985, Vol. 13, No. 13 4699-4717
© 1985

Articles

Construction of cDNA coding for human von Willebrand factor using antibody probes for colony-screening and mapping of the chromosomal gene

Cornelis L. Verweij, Carlie J.M. de Vries, Ben Distel, Anton-Jan van Zonneveld, Ad Geurts van Kessel+, Jan A. van Mourik* and Hans Pannekoek§

Department of Molecular Biology, Central Laboratory of the Netherlands Red Cross Blood Transfusion Service P.O. Box 9406, 1006 AK Amsterdam *Department of Blood Coagulation, Central Laboratory of the Netherlands Red Cross Blood Transfusion Service P.O. Box 9406, 1006 AK Amsterdam +Department of Cell Biology and Genetics, Erasmus University Rotterdam, The Netherlands

§To whom correspondence should be addressed

Received April 15, 1985. Revised June 11, 1985. Accepted June 18, 1985.

Von Willebrand Factor (vWF) mRNA was identified in fractionated polyA+ RNA preparations isolated from cultured human endothelial cells. Micro-injection of specific polyA+ RNA fractions in Xenopus laevis oocytes provoked the synthesis of a vWF-like product which could be detected with an imunoradiometric assay relying on Sepharose-linked monoclonal anti-vWF IgG and different radiolabeled monoclonal anti-vWF IgGs. A vWF-mRNA-containing polyA+ RNA preparation served as substrate for a size-selected cDNA-expression library of 60 000 colonies which was screened for the synthesis of antigens related to vWF, using polyclonal anti-vWF IgG and a second antibody conjugated with peroxidase. Eight positive colonies were detected of which two reacted strongly in the enzyme-linked assay. Immunoblotting of bacterial extracts of "expression clones" with a monoclonal anti-vWF IgG revealed polypeptides which size fits within the length of the cDNA insertions. Northern blotting of human endothelial RNA, employing fragments of vWF cDNA as probes, showed specific hybridization with a mRNA of about 9000 nucleotides. DNA-sequence analysis of a vWF-cDNA insertion revealed an open reading frame followed by a translation stopcodon. It is argued that the cDNA insertions encode the carboxy-terminal part of the vWF protein. vWF-cDNA probes were employed to map the von Willebrsnd factor gene on chromosome 12 using a panel of 35 human-rodent somatic cell hybrids.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 BloodHome page
P. D. James, L. A. O'Brien, C. A. Hegadorn, C. R. P. Notley, G. D. Sinclair, C. Hough, M.-C. Poon, and D. Lillicrap
A novel type 2A von Willebrand factor mutation located at the last nucleotide of exon 26 (3538G>A) causes skipping of 2 nonadjacent exons
Blood, November 1, 2004; 104(9): 2739 - 2745.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
C. P. M. Hayward, E. M. Cramer, Z. Song, S. Zheng, R. Fung, J.-M. Masse, R. H. Stead, and T. J. Podor
Studies of Multimerin in Human Endothelial Cells
Blood, February 15, 1998; 91(4): 1304 - 1317.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.