Cloning and Expression of CD19, a Human B-Cell Marker in NIH-3T3 Cell Line.

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 DH5α competent bacteria. After blue/white selection, one positive white colony was subjected to plasmid extraction and direct sequen

SUBMITTER: Abbasi-Kenarsari H 

PROVIDER: S-EPMC4388889 | biostudies-literature | 2015 Jan-Mar

REPOSITORIES: biostudies-literature