Difference between revisions of "Part:BBa K3573002:Experience"

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===Applications of BBa_K3573002===
 
===Applications of BBa_K3573002===
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Our team has amplified HRAS using PCR and cloned the gene into an expression vector.
  
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Korea_HS team have performed binding assays using size-exclusion chromatography. We ran three separate runs of 3 superdex75 10/300GL column with buffer (20mM Tris-HCl pH 7.0+250mM NaCl)
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[[File:T--Korea_HS--RasPCR.jpg|500px]]
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1. scFv(RAS) only
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2. HRAS only
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3. scFv(RAS)+HRAS
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The result showed that HRAS's elution peak shifted to higher molecular weight position in the presence of scFv(RAS) compared to HRAS only, indicating their binding.
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Plates obtained from transformation
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[[File:T--Korea_HS--RASPlates.jpg|500px]]
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We have tested protein expression in 4 different conditions (2 temperatures X 2 IPTG concentrations). After expression, Ni-NTA column was used for protein purification using the His6-tag located at the N-terminus of CPP-scFv(RAS).
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[[File:T--Korea_HS--RasNiNTA.jpg|800px]]
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 00:17, 26 October 2020


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K3573002

Our team has amplified HRAS using PCR and cloned the gene into an expression vector.

T--Korea HS--RasPCR.jpg

Plates obtained from transformation

T--Korea HS--RASPlates.jpg


We have tested protein expression in 4 different conditions (2 temperatures X 2 IPTG concentrations). After expression, Ni-NTA column was used for protein purification using the His6-tag located at the N-terminus of CPP-scFv(RAS).

T--Korea HS--RasNiNTA.jpg

User Reviews

UNIQ483f3357c600ac33-partinfo-00000000-QINU UNIQ483f3357c600ac33-partinfo-00000001-QINU