[Chicken SREBP1 gene antiserums preparation and its tissue expression characterization]

Area of Science:

• Molecular Biology
• Animal Science
• Biochemistry

Background:

• Sterol regulatory element binding protein 1 (SREBP1) plays a crucial role in lipid metabolism and adipogenesis.
• Understanding SREBP1 expression patterns in poultry is vital for optimizing meat production and quality.
• Chicken SREBP1's specific functions and tissue distribution require detailed investigation.

Purpose of the Study:

• To generate high-titer and specific antibodies against chicken sterol regulatory element binding protein 1 (SREBP1).
• To investigate the tissue-specific expression profile of SREBP1 in chickens.
• To compare SREBP1 expression levels between fat and lean broiler lines.

Main Methods:

• Analysis of chicken SREBP1 nuclear import sequence to identify antigen epitopes.
• Amplification and cloning of the SREBP1 epitope-coding fragment into an expression vector.
• Recombinant protein expression in E. coli, purification via affinity chromatography, and antibody production in rabbits.
• Detection of antibody titer and specificity using ELISA and Western blot; tissue expression analysis.

Main Results:

• Successfully prepared chicken SREBP1 antibodies with a high titer (1:102,400) and specificity.
• SREBP1 expression was significantly higher in abdominal adipose tissue and heart, with lower levels in liver, muscle, stomach, duodenum, and spleen.
• No SREBP1 signal was detected in breast and leg muscle; significantly higher expression in the abdominal adipose tissue of fat broiler lines compared to lean lines (P<0.05).

Conclusions:

• The developed antiserum against chicken SREBP1 is highly specific and effective.
• SREBP1 is predominantly expressed in the abdominal adipose tissue of broilers, indicating its importance in fat deposition.
• Fat broiler lines exhibit higher SREBP1 expression than lean lines, suggesting a genetic basis for fat accumulation.