An improved strategy for tandem affinity purification-tagging of Schizosaccharomyces pombe genes

作者：Cipak Lubos; Spirek Mario; Novatchkova Maria; Chen Zhiming; Rumpf Cornelia; Lugmayr Wolfgang; Mechtler Karl; Ammerer Gustav; Csaszar Edina; Gregan Juraj^*

来源：Proteomics, 2009, 9(20): 4825-4828.

DOI：10.1002/pmic.200800948

• MS
• Schizosaccharomyces pombe
• Tandem affinity purification
• Technology

Tandem affinity purification (TAP) is a method that allows rapid purification of native protein complexes. We developed an improved technique to fuse the fission yeast genes with a TAP tag. Our technique is based on tagging constructs that contain regions homologous to the target gene cloned into vectors carrying a TAP tag. We used this technique to design strategies for TAP-tagging of predicted Schizosaccharomyces pombe genes (http://mendel.imp.ac.at/Pombe_tagging/). To validate the approach, we purified the proteins, which associated with two evolutionarily conserved proteins Swi5 and Sfr1 as well as three protein kinases Ksg1, Orb6 and Sid1.

• 出版日期2009-10
• 单位吉林大学