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RTG-P1

規格:

價格:

編號:B215542

品牌:Mingzhoubio

標準菌株

定量菌液

DNA

RNA

規格:

凍干粉

斜面

甘油

平板

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產品名稱

RTG-P1

商品貨號

B215542

Organism

Oncorhynchus mykiss, trout, rainbow

Tissue

mixed; testis; ovary

培養基

Product Format

frozen

Morphology

fibroblast

Culture Properties

adherent

Biosafety Level

2 Cells contain SV-40 and CMV viral DNA sequences

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Gender

male and female mixed

Applications

The vector, pGL3-prMx1-Basic-Neo, expresses the firefly luciferase gene under the control of the promoter for the IFN-induced gene Mx1. The vector contains SV40 viral DNA sequences and the neomycin resistance gene.

This cell line can be used to study the activation of the IFN pathway.

The RTG-2 cell line (ATCC CCL-55) was transfected with a trout Mx1 promoter-luciferase construct and selected to produce a stable cell line, RTG-P1 [PubMed: 15043947].

The RTG-P1 reporter system constitutes an interesting tool to study the induction and regulation of IFN signaling in teleost fish.

Storage Conditions

liquid nitrogen vapor phase

Images

Clinical Data

male and female mixed

Comments

The RTG-2 cell line (ATCC CCL-55) was transfected with a trout Mx1 promoter-luciferase construct and selected to produce a stable cell line, RTG-P1 [PubMed: 15043947]. The vector, pGL3-prMx1-Basic-Neo, expresses the firefly luciferase gene under the control of the promoter for the IFN-induced gene Mx1. The vector contains SV40 viral DNA sequences and the neomycin resistance gene. Luciferase expression is stable and highly induced by polyinosinic:polycytidylic acid (poly I:C) in RTG-P1 cells [PubMed: 15043947]. This cell line can be used to study the activation of the IFN pathway. The RTG-P1 reporter system constitutes an interesting tool to study the induction and regulation of IFN signaling in teleost fish.

Complete Growth Medium

Leibovitz's L-15 medium with 2 mM L-glutamine supplemented with 0.2 mg/ml G418 and 10% fetal bovine serum.

(Note: The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation)

Subculturing

Protocol:

Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
An inoculum of 6 X 10(3) to 1 X 10(4) viable cells/sq. cm is recommended.
Incubate cultures at 22C.

Interval: Maintain cultures at a cell concentration between 1 X 10(4) and 1 X 10(5) cells/sq. cm

Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended

Medium Renewal: Two to three times weekly

Cryopreservation

Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

Culture Conditions

Atmosphere: air, 100%

Temperature: 22.0°C

Population Doubling Time

approximately 67 hours

Name of Depositor

B Collet

References

Collet B, et al. An Mx1 promoter-reporter system to study interferon pathways in rainbow trout. Dev. Comp. Immunol. 28: 793-801, 2004. PubMed: 15043947

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