Order this document
Anton Yuryev: I still consider the vectors described in this paper superior to the 2mn vectors for the two-hybrid. Segregation efficiency of CEN based plasmid is about 95% and 2mn vectors is only 40%. Consider yeast cell that just was transformed with two plasmids. The probability of loosing any of it is about 56%. Because the loss of any plasmid leads to cell death, your chances to loose important interaction is above 50% !
Proc Natl Acad Sci U S A 1992 Jul 1;89(13):5789-93
To identify proteins that interact with Jun or Fos we have used the protein interaction cloning system developed by S. Fields and O.-K. Song [(1989) Nature (London) 340, 245-246] to clone mammalian cDNAs encoding polypeptides that interact with the dimerization and DNA-binding motif (bZIP; basic domain leucine zipper motif) of Jun. For this purpose, yeast cells lacking GAL4 activity but expressing a GAL4 DNA-binding domain-Jun bZIP fusion protein were transformed with a mouse embryo cDNA plasmid library in which the cDNA was joined to a gene segment encoding the GAL4 transcriptional activation domain. Several transformants exhibiting GAL4 activity were identified and shown to harbor plasmids encoding polypeptides predicted to form coiled-coil structures with Jun and/or Fos. One of these is a bZIP protein of the ATF/CREB protein family--probably the murine homolog of TAXREB67. Two others encode polypeptides with predicted potential to form coiled-coil structures, and seven other isolates encode segments of alpha- or beta-tropomyosin, classical coiled-coil proteins. The tropomyosin polypeptides were found to interact in the yeast assay system with the bZIP region of Jun but not with the bZIP region of Fos. Our results illustrate the range of protein interaction cloning for discovering proteins that bind to a given target polypeptide.
PMID: 1631061, UI: 92335183
the above report in MacintoshPCUNIXTextHTMLformat documents on this page through Loansome Doc