Cloning, sequencing, and analysis of cDNA encoding bovine granulocyte-colony stimulating factor

Area of Science:

• Immunology
• Molecular Biology
• Biochemistry

Background:

• Neutrophils are crucial for combating bacterial infections.
• Granulocyte colony-stimulating factor (G-CSF) promotes neutrophil development and function.
• G-CSF enhances neutrophil phagocytosis and pathogen killing.

Purpose of the Study:

• To isolate and sequence the cDNA encoding bovine G-CSF (bG-CSF).
• To analyze the sequence identity of bG-CSF compared to other species.
• To investigate G-CSF mRNA expression in bovine immune cells.

Main Methods:

• cDNA library screening using primers from ovine G-CSF.
• Full-length cDNA sequencing and open reading frame analysis.
• Sequence alignment and identity comparison with other species' G-CSF.
• Reverse transcription polymerase chain reaction (RT-PCR) for mRNA expression analysis.

Main Results:

• A 1460-nucleotide cDNA clone for bG-CSF was isolated.
• The bG-CSF sequence exhibits high identity to ovine (95%), porcine (89%), human (85%), and murine (76%) G-CSF.
• The deduced bG-CSF protein (174 amino acids) shares high identity with orthologs.
• Bovine neutrophils do not express G-CSF mRNA, but mononuclear cells do after 48h incubation.

Conclusions:

• Bovine G-CSF cDNA has been successfully isolated and sequenced.
• The high sequence conservation suggests functional similarity across species.
• G-CSF expression is primarily associated with mononuclear cells in cattle, not neutrophils.