Mobile website
Expression of Mutant M-CSF from Human Leukemic Cell Line J6-1 and the Binding Activity for Its Receptor
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective: To clone and express functional part of mutant M-CSF (muM-CSF) from human leukemic cell line J6-1 and investigate its Kd for dissociation and biological activity on the proliferation of J6-1.Method: Functional part of muM-CSF was cloned by RT-PCR and inserted into pET32c(+) and expressed in E.coli BL21trxB (DE3). The recombinant protein was purified through Ni2+ affinity column and antibody linked affinity column. ELISA was performed to define the Kd of the muM-CSF to its receptor. Colony formation assay was performed to test its effects on the proliferation of J6-1. Results: The protein was purified and its Kd to the receptor was 3.7 nmol/L. muM-CSF showed elevated proliferation-stimulating potential than normal M-CSF. Conclusion: muM-CSF could be expressed in the pET32c(+), BL21 system and the muM-CSF showed elevated proliferation promoting ability as to normal M-CSF.
Keywords:
Project Supported:
Clc Number:
Q786
