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pVITRO2-Lucia/SEAP

pVITRO2-hygro-Lucia/SEAP Unit size Cat. code Docs Qty Price
Lucia/SEAP - Hygromycin resistance
20 µg
pvitro2-lucsp
+-
$433.00
pVITRO2-blasti-Lucia/SEAP Unit size Cat. code Docs Qty Price
Lucia/SEAP - Blasticidin resistance
20 µg
pvitro2-blucsp
+-
$433.00
pVITRO2-neo-Lucia/SEAP Unit size Cat. code Docs Qty Price
Lucia/SEAP - Neomycin resistance
20 µg
pvitro2-nlucsp
+-
$433.00

Expression plasmids pVITRO2-Lucia/SEAP are developed mainly for in vitro studies.
pVITRO2-Lucia/SEAP allows the ubiquitous and constitutive co-expression of two reporter genes: Lucia (secreted luciferase) and SEAP (secreted embryonic alkaline phosphatase). pVITRO2-Lucia/SEAP plasmids can be stably transfected in mammalian cells and the reporter genes are expressed at high levels.

pVITRO2-Lucia/SEAP plasmids carry two human ferritin composite promoters, FerH (heavy chain) and FerL (light chain). To eliminate the iron regulation, their 5’UTRs have been replaced by the 5’UTR of the mouse and chimpanzee EF-1α genes. pVITRO2-Lucia/SEAP plasmids are available with different selectable markers that are active both in E. coli and mammalian cells.

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Specifications

Promoters: SV40/hFerH/mEF1α & CMV/hFerL/chEF1α
Selection markers active both in E. coli and mammalian cells:
  - bsr (blasticidin resistance)
  - hph (hygromycin resistance)
  - neo (kanamycin/G418 resistance).
Reporter Genes:
  - Lucia luciferase (secreted luciferase)
  - SEAP (secreted embryonic alkaline phosphatase)

These products are covered by a Limited Use License (See Terms and Conditions).

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Contents

- 20 µg of lyophilized DNA
- 4 pouches of the appropriate E. coli Fast-Media® (2 TB and 2 Agar)

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Description

Gene Combinations in a Single Plasmid

Co-expression of two or more genes from a single vector is more        efficient and convenient than using two separate vectors.   pVITRO2-Lucia/SEAP      are multigenic plasmids that contain two distinct   transcription  units     (TU).

Strong and Constitutive Expression of Two Transgenes

Each pVITRO2-Lucia/SEAP plasmid features two constitutive promoters   that   drive high levels of expression from two separate TU in a large   number   of mammalian cell lines.
Transcriptional interference is   minimized   by using promoters that are coordinately activated, and   strong   polyadenylation signals (polyA). pVITRO2-Lucia/SEAP plasmids   contain two  human  ferritin composite promoters, FerH (heavy chain) and   FerL (light   chain), combined to the SV40 and CMV enhancers   respectively. To   eliminate the iron regulation, their 5’UTRs have been   replaced by the   5’UTR of the mouse and chimpanzee EF-1α genes.

Single Selection Marker for E. coli and Mammalian Cells

pVITRO2-Lucia/SEAP plasmids are available with a selectable marker that is active both in E. coli and mammalian cells: bsr (blasticidin resistance), hph (hygromycin resistance) or neo (kanamycin/G418 resistance).
In bacteria, the resistance gene is expressed from the E. coli EM7 promoter. In mammalian cells, it is transcribed from the promoter      located 3’ of the Ori, as a polycistronic mRNA and translated through      the IRES of the Foot and Mouth Disease Virus.

Two Reporter Genes

pVITRO2-Lucia/SEAP express two reporter genes: Lucia luciferase and SEAP
Lucia secreted luciferase and SEAP (secreted embryonic alkaline phosphatase) are commonly used reporter proteins.
Lucia is a secreted   luciferase with strong bioluminescent activity. The Lucia luciferase reporter  gene  is ideal for promoter activity and gene expression studies. Lucia luciferase activity can be detected and quantified directly in the   culture medium of transfected cells using QUANTI-Luc™ detection reagent.

pVITRO2-Lucia/SEAP plasmids can be used as control plasmids for pVITRO2-MCS or cloning   vectors. The reporter genes are flanked by unique    restriction sites and   can be easily replaced by open reading frames    chosen from InvivoGen’s Gene A-List.

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Details

pVITRO2-Lucia/SEAP Map• hFerH and hFerL composite promoters: Ferritin is a    24 subunit protein composed of two subunit types, termed H (heavy)  and  L  (light). Ferritin is ubiquitously expressed. Its synthesis is  highly   regulated by the iron status of the cell. To eliminate the iron    regulation of the ferritin promoters, the 5’UTR of FerH and FerL have    been replaced by the 5’UTR of the mouse and chimpanzee elongation  factor   1 (EF1) genes, respectively.

• SV40 enhancer which is comprised of a 72-base-pair    repeat allows the enhancement of gene expression in a large host   range.  The enhancement varies from 2-fold in non-permissive cells to   20-fold  in permissive cells.

• CMV enhancer: The major immediate early enhancer   of the human cytomegalovirus  (HCMV) is composed of unique and repeated   sequence motifs. The HCMV  enhancer can substitute for the 72-bp  repeats  of SV40 and is severalfold  more active than the SV40 enhancer.

• SV40 pAn: the Simian Virus 40 late polyadenylation    signal enables efficient cleavage and polyadenylation reactions    resulting in high levels of steady-state mRNA.

• pMB1 ori: a minimal E. coli origin of replication to limit vector size, but with the same activity as the longer Ori.

• FMDV IRES: The internal ribosome entry site of the    Foot and Mouth Disease Virus enables the translation of two open    reading frames from one mRNA with high levels of expression.

• EM7 is a bacterial promoter that enables the constitutive expression of the antibiotic resistance gene in E. coli.

•  Antibiotic resistance gene :
- Resistance to Blasticidin S is conferred by the bsr gene from Bacillus cereus.
- hph gene confers resistance to Hygromycin B both in E. coli and mammalin cells.
- The neo gene from Tn5 confers resistance to Kanamycin in E.coli and G418 in mammalian cells.
In bacteria, the resistance gene is expressed from the constitutive E.     coli EM7  promoter. In mammalian cells, the resistance gene is     transcribed from the rat EF-1α  promoter as a polycistronic mRNA and     translated via the FMDV IRES.

• EF1 pAn is a strong polyadenylation signal.

• Lucia luciferase gene: Lucia is a secreted  luciferase with strong bioluminescent activity.

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