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Step-tag expression system

Step-tag expression system. pASK-IBA VECTOR. Introduction. pASK-IBA VECTOR. Properties Tet-promoter Strep-tag2 Protease cleavage site Periplasm secretion sequence. Tet-off system. Tet repressor binding domain + V16 activator domain. Tet-on system.

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Step-tag expression system

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  1. Step-tag expression system pASK-IBA VECTOR

  2. Introduction

  3. pASK-IBA VECTOR • Properties • Tet-promoter • Strep-tag2 • Protease cleavage site • Periplasm secretion sequence

  4. Tet-off system Tet repressor binding domain + V16 activator domain

  5. Tet-on system Tet repressor binding domain + V16 activator domain, change 4 a.a

  6. TREX system

  7. TREX system

  8. Strep-tag2 • Advantage of strep2 tag • does not interfere with folding or bioactivity • one of the most stable proteins • High affinity(derivated biotin) Trp-Ser-His-Pro-Gln-Phe-Glu-Lys

  9. Various cleavage, secretion site 1.Xa, thrombin, TEV protease site 2.Possible secretion to periplasm

  10. Advantages of periplasm secretion • Soluble form • Easy to purify • Stability(protease et) • Decrese of unfolding form

  11. purification regeneration loading

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