Citation
- Authors: Abbasi-Kenarsari, H., Shafaghat, F., Baradaran, B., Movassaghpour, A. A., Shanehbandi, D., Kazemi, T.
- Year: 2015
- Journal: Avicenna J Med Biotechnol
- Applications: in vitro / DNA / jetPEI
- Cell type: NIH/3T3
Description: Murine embryonic fibroblasts
Known as: NIH/3T3, 3T3
Abstract
BACKGROUND:
CD19 is a pan B cell marker that is recognized as an attractive target for antibody-based therapy of B-cell disorders including autoimmune disease and hematological malignancies. The object of this study was to stably express the human CD19 antigen in the murine NIH-3T3 cell line aimed to be used as an immunogen in our future study.
METHODS:
Total RNA was extracted from Raji cells in which high expression of CD19 was confirmed by flow cytometry. Synthesized cDNA was used for CD19 gene amplification by conventional PCR method using Pfu DNA polymerase. PCR product was ligated to pGEM-T Easy vector and ligation mixture was transformed to DH5