Estimated Removal of Live Colony date: 26 March 2020.Peritoneal neutrophils and macrophages, bone marrow cells and neutrophils isolated from bone marrow of mice homozygous for Ncf1m1J spontaneous mutation fail to produce superoxide upon stimulation in vitro with N-formyl-methionyl-leucyl-phenylalanine (fMLP) or phorbol 12-myristate 13-acetate (PMA). Read More +
Peritoneal neutrophils and macrophages, bone marrow cells and neutrophils isolated from bone marrow of mice homozygous for Ncf1m1J fail to produce superoxide upon stimulation in vitro with N-formyl-methionyl-leucyl-phenylalanine (fMLP) or phorbol 12-myristate 13-acetate (PMA), as determined by kinetic spectrophotometric analysis of cytochrome c reduction. Western blot analysis detected no full-length NCF1/p47phox protein in cells from these mice; a faint band of slightly smaller molecular size than the wild type NCF1 protein was observed on probing with antibodies to NCF1. To exclude the possibility that the NCF1 protein is produced in cells of mutant mice but is degraded rapidly by endogenous proteases, bone marrow cells were isolated and samples prepared for western blot analysis in the presence of diisopropyl fluorophosphate (DFP); no difference was observed upon analysis of freshly prepared cell lysates made with and without DFP, indicating that NCF1 protein is not expressed in mutant cells (Huang et al. 2000).
Analysis for other NADPH oxidases involved in neutrophil superoxide production revealed that NCF2/p67phox was present at wild type levels and CYBB/gp91phox and CYBA/p22phox were expressed at higher than wild type levels. Other neutrophil products were assayed and found not to differ in bone marrow cells of C57BL/6J vs. mutant mice: GR-1, a granulocyte marker present on approximately 40% of marrow cells by fluorescence activated cell sorting (FACS) analysis; NCF4/p40phox and RAC1/RAC2 proteins, analyzed by western blot analysis; and expression of mRNAs encoding the granule proteins myeloperoxidase, elastase, cathepsin, lysozyme, lactoferrin and gelatinase as detected by reverse transcriptase - polymerase chain reaction (RT-PCR) (Huang et al. 2000).
The response of Ncf1m1J/Ncf1m1J mice to infectious or irritant (inflammatory stimulus) challenge has not been investigated (Huang et al. 2000). However, mice homozygous for the targeted mutation Ncf1tm1Shl (Stock No. 027331) - which disrupts the gene near the point of the Ncf1m1J mutation and, like the latter, results in failure of phagocytes to generate a superoxide burst following PMA stimulation in vitro - developed spontaneous infections with a variety of microorganisms including Staphylococcus xylosus, Lactobacillus, a hyaline septate mold and the fungus Paecilomyces, despite being maintained under specific pathogen free conditions. Ten-fold more Staphyloccoccus aureus bacteria survived intracellularly following in vitro uptake by phagocytes inblood from these mice than from control littermates, demonstrating a defect in the intracellular bactericidal activity of the mutant mice. Following intraperitoneal injection of the sterile irritant thioglycolate, twice as many leukocytes were recruited to the peritoneal cavities of Ncf1tm1Shl homozygous mice than of littermate controls; proportions of neutrophils in mutant and control exudates were similar (Jackson et al., J Exp Med 182:751-758 1995).
This spontaneous mutation arose in the B6.BKS(D)-Leprdb/J (Stock No. 000697) production colony at The Jackson Laboratory. The mutation was identified in 1999. (Huang CK. et al. 2000) The mutations Leprdb and m have been selectively bred out of this strain by crossing to C57BL/6J at least three times.
|Allele Name||mutation 1, Jackson|
|Allele Synonym(s)||Ncf1*; p47-; p47phox; p47phox-`; rs230824082|
|Gene Symbol and Name||Ncf1, neutrophil cytosolic factor 1|
|Strain of Origin||B6.Cg-Dock7m +/+ Leprdb/J|
|Molecular Note||The mutation is an A-to-C transversion at the -2 position 5' off exon 8 in intron 7, changing splice acceptor site CAG to CCG. This results in aberrant splicing of the transcript. Immunoblotting detected no intact NCF1 protein in cells from these mice.|
When using the B6(Cg)-Ncf1m1J/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #004742 in your Materials and Methods section.
"MICE" means mouse strains, their progeny derived by inbreeding or crossbreeding, unmodified derivatives from mouse strains or their progeny supplied by The Jackson Laboratory ("JACKSON"). "PRODUCT(S)" means biological materials supplied by JACKSON, and their derivatives. "SERVICES" means projects conducted by JACKSON for other parties that may include but are not limited to the use of MICE or PRODUCTS. "RECIPIENT" means each recipient of MICE, PRODUCTS, or SERVICES provided by JACKSON including each institution, its employees and other researchers under its control. MICE or PRODUCTS shall not be: (i) used for any purpose other than internal research, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services. Acceptance of MICE, PRODUCTS or SERVICES from JACKSON shall be deemed as agreement by RECIPIENT to these conditions, and departure from these conditions requires JACKSON’s prior written authorization.
MICE, PRODUCTS AND SERVICES ARE PROVIDED "AS IS". JACKSON EXTENDS NO WARRANTIES OF ANY KIND, EITHER EXPRESS, IMPLIED, OR STATUTORY, WITH RESPECT TO MICE, PRODUCTS OR SERVICES, INCLUDING ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, OR ANY WARRANTY OF NON-INFRINGEMENT OF ANY PATENT, TRADEMARK, OR OTHER INTELLECTUAL PROPERTY RIGHTS.
In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of, PRODUCTS or SERVICES, JACKSON will, at its option, provide credit or replacement for the PRODUCT received or the SERVICES provided; JACKSON makes no other representations and this shall be the exclusive remedy of the purchaser. Please note specific policy for live mice.
Consistent with the requirement for a written understanding regarding animal care and use, the JACKSON Animal Care and Use Committee will review the animal care and use protocol(s) associated with any SERVICES to be performed at JACKSON, and JACKSON shall have ultimate responsibility and authority for the care of animals while on site or in JACKSON custody.
In no event shall JACKSON, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, PRODUCTS, or SERVICES, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of JACKSON, its agents or employees. Unless prohibited by law, in purchasing or receiving MICE, PRODUCTS, or SERVICES from JACKSON, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges JACKSON from all such causes of action or damages, and further agrees to defend and indemnify JACKSON from any costs or damages arising out of any third party claims.
MICE, PRODUCTS or SERVICES are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.
The foregoing represents the General Terms and Conditions applicable to JACKSON’s MICE, PRODUCTS or SERVICES. In addition, special terms and conditions of sale of certain MICE, PRODUCTS, or SERVICES may be set forth separately in JACKSON web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, PRODUCTS and SERVICES by JACKSON, and by its licensees and distributors.
Acceptance of delivery of MICE, PRODUCTS or SERVICES shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on JACKSON, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, PRODUCTS or SERVICES by JACKSON.