Abstract
Yeast two-hybrid approach is popularly used nowadays as an important technical method in the field of studying protein-protein interactions. Although yeast two-hybrid system is obviously advantageous in searching interacting proteins and setting up the network of proteins interaction, not all of proteins can use routine yeast two-hybrid method to search interacting proteins. Many important proteins, such as some nucleoprotein transcriptional factor, carry out the regular method and construct the bait-BD vector to screen the library containing AD vector. However, it usually results in failures because it contains the activate domain and can self-activate the reporter gene. In this study, we changed the research strategy, fused the bait gene (FOXA3) with the AD vector to screen the library containing BD vector, so that we constructed a two-hybrid library containing BD vector and can bypass the interference of self-activation. And we used this two-hybrid library to screen FOXA3, a hepatocyte nuclear factor, and found out an interacting protein: complement component C3.
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Ang S.-L., Wierda A., Wong D., Stevens K.-A., Cascio S., Rossant J., Zaret K.-S., The formation and maintenance of the definitive endoderm lineage in the mouse: involvement of HNF3/forkhead proteins. Development, 1993, 119(6):1301–1315
Blackstock S.-G. and Weir M.-P., Proteomics: quantitative and physical mapping of cellular proteins, Trends Biotechnol., 1999, 17(3): 121–127
Cianflone K., Zakarian R. and Couillard C., Fasting acylation-stimulating protein is predictive of postprandial triglyceride clearance, J. Lipid. Res., 2004, 45(1): 124–131
Faraj M., Havel P.-J. and Phelis S., Plasma acylation-stimulating protein, adiponectin, leptin, and ghrelin before and after weight loss induced by gastric bypass surgery in morbidly obese subjects, J. Clin. Endocrinol. Metab., 2003, 88(4): 1594–1602
Fong K.-Y., Botto M. and Walport M.-J., Genomic organization of human complement component C3, Genomics, 1990, 7(4): 579–586
Gietz D., St Jean A., Woods R.-A., Schiestl P.-H., Improved method for high efficiency transformation of intact yeast cells, Nucleic. Acids. Res., 1992, 20(6): 1425
Inoue H., Nojima H. and Okayama H., High efficiency transformation of Escherichia coli with plasmids, Gene, 1990, 96(1): 23–28
Katoh M. and Katoh M., Human FOX gene family, Int. J. Oncol., 2004, 25(5):1495–1500
Lantz K.-A. and Kaestner K.-H., Winged-helix transcription factors and pancreatic development, Clin. Sci., 2005, 108(3): 195–204
Monaghan A.-P., Kaestner K.-H., Grau E., Schutz G., Postimplantation expression patterns indicate a role for the mouse forkhead/HNF-3, and genes in determination of the definitive endoderm, chordamesoderm and neuroectoderm, Development, 1993, 119(3): 567–578
Odom D.-T., Zizlsperger N., Gordon D.-B., Gordon D.-B., Bell G.-W., Dinaldi N.-J., Murray H.-L., Volkert T.-L., Schreiber J., Rolfe P.-A., Gifford D.-K., Fraenkel E., Bell G.I., Young R.-A., Control of pancreas and liver gene expression by HNF transcription factors, Science, 2004, 303(5662): 1378–1381
Shen W., Scearce L.-M., Brestelli J.-E., Foxa3 (hepatocyte nuclear factor 3γ)is required for the regulation ofhepatic GLUT2 expression and the maintenance of glucose homeostasis during a prolonged fast, J. Boil. Chem., 2001, 276(46): 42812–42817
Ylitalo K., Nuotio I. and Viikari J., C3, hormone-sensitive lipase, and peroxisome proliferator-activated receptor gamma expression in adipose tissue of familial combined hyperlipidemia patients, Metabolism, 2002, 51(5): 664–670
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Translated from Journal of Fudan Universrty (Natural Science), 2005, 44(4)[译自: 复旦学报(自然科学版),2005,44(4)]
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Wan, B., Shi, Y. & Huo, K. An improved yeast two-hybrid approach for detection of interacting proteins. Front. Biol. China 1, 120–126 (2006). https://doi.org/10.1007/s11515-006-0008-9
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DOI: https://doi.org/10.1007/s11515-006-0008-9