Human ROM1 cDNA Clone in Bacterial Expression Vector (His-GST)
cDNA
Clon
Bacterial Expression Vector
pPB-His-GST
Kanamycin
T7 Promoter
His-GST
Transient
Catalog No. ABIN4839240
Quick Overview for Human ROM1 cDNA Clone in Bacterial Expression Vector (His-GST) (ABIN4839240)
Gene
ROM1
(Retinal Outer Segment Membrane Protein 1 (ROM1))
Fusion tag
His-GST
Application
Cloning (Clon)
Insert
cDNA
Vector
Bacterial Expression Vector
Vector Backbone
pPB-His-GST
Promoter
T7 Promoter
Bacterial Resistance
Kanamycin
Expression Type
Transient
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Species
- Human
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Purpose
- Bacterial expression of Human ROM1 with His-GST
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Specificity
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5-NheI and 3-XhoI
Fusion tag: Dual N-terminal tag, 6X Histidine followed by Glutathione-S-Transferase Protein which is cleavable with TEV (Size 27.9 kDa) -
Insert Length
- 1056 bp
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Sequencing Primer
- GST Forward primer: 5'-CACGTTTGGTGGTGGCGAC3', T7 terminator primer: 5'-GCTAGTTATTGCTCAGCGG-3'
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Application Notes
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The pPB vectors are low-medium copy number vectors in which the gene expression is driven by the strong T7 promoter.
Below are some basic guidelines for using the pPB vectors for protein production:
1. The pPB vectors are designed to be used with E. coli strains that are DE3 lysogens i.e. the host E. coli cell has a source of T7 RNA polymerase.
2. Recombinant protein induction is usually done at OD600 of 0.6-1.2 using Isopropyl β-D-1-thiogalactopyranoside (IPTG) at a final concentration of 0.05 -1mM.
3. The ideal concentration of IPTG must be determined empirically for each recombinant protein/cell-line. Similarly, the length of time and temperature for induction provide other variables that need to be optimized on a case-to-case basis.
4. For toxic proteins, it is recommended to go for shorter induction time and also to try and suppress basal recombinant gene expression through (a) addition of glucose or use of pLysS plasmid. Please note that special cell-lines are also available in the market that cater to expression of toxic proteins.
5. Once grown for the desired length of time, harvest cells by centrifugation and either freeze the cells at -80°C (as such or after re-suspending in the desired buffer) or proceed with the purification. -
Restrictions
- For Research Use only
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Format
- Liquid
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Buffer
- 10 mM Tris-HCI, 1 mM EDTA, pH 8.0
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Storage
- -20 °C
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Storage Comment
- 1 year when stored at -20° C or lower in a non-frost free freezer.
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Expiry Date
- 12 months
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- ROM1 (Retinal Outer Segment Membrane Protein 1 (ROM1))
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Alternative Name
- ROM1
Target
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