Random PCR mutagenesis screening of secreted proteins by direct expression in mammalian cells.
نویسندگان
چکیده
منابع مشابه
Random PCR mutagenesis screening of secreted proteins by direct expression in mammalian cells.
We have developed a general method for screening randomly mutagenized expression libraries in mammalian cells by using fluorescence-activated cell sorting (FACS). The cDNA sequence of a secreted protein is randomly mutagenized by PCR under conditions of reduced Taq polymerase fidelity. The mutated DNA is inserted into an expression vector encoding the membrane glycophospholipid anchor sequence ...
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We thank John Connolly for helpful discussions and critical reading of the manuscript. We are very grateful to Fabrice Chareyre, Laetitia Gressin, Catherine Giudicelli, Jean Christophe Beaudoin and Isabelle Legall for help in the BAC screening, sequencing and genotyping. This work was supported by a grant from the Association pour la Recherche sur le Cancer (ARC) and the Ministère de l’Educatio...
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ژورنال
عنوان ژورنال: Proceedings of the National Academy of Sciences
سال: 1992
ISSN: 0027-8424,1091-6490
DOI: 10.1073/pnas.89.12.5467