High-level expression of biologically active chicken prolactin in E. coli

Area of Science:

• Biotechnology
• Molecular Biology
• Endocrinology

Background:

• Chicken prolactin (cPRL) is a crucial avian hormone.
• Efficient production of recombinant cPRL is essential for research and potential applications.

Purpose of the Study:

• To develop a method for large-scale production of biologically active recombinant chicken prolactin (cPRL) in E. coli.
• To characterize the immunological and biological properties of the produced recombinant cPRL.

Main Methods:

• Genetic engineering of cPRL cDNA and expression vector pKK223-3 for E. coli expression.
• Insertion of a DNA linker with Shine-Dalgarno sequences to enhance protein production.
• Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for molecular mass determination.
• Immunological binding assays using antiserum against turkey prolactin.
• In vivo biological assays measuring pigeon crop sac mucosal weight.

Main Results:

• A significant quantity of recombinant cPRL was produced in E. coli.
• SDS-PAGE confirmed the recombinant cPRL protein has a molecular mass of 23 kDa.
• The recombinant cPRL exhibited equivalent binding kinetics to an antiserum for turkey prolactin.
• Biological activity was confirmed by comparable pigeon crop sac mucosal weight increase as turkey prolactin.

Conclusions:

• The engineered E. coli system effectively produced recombinant cPRL.
• The recombinant cPRL possesses identical immunological and biological functions to authentic avian prolactin.
• This provides a viable source for studying avian prolactin functions.