Isolation and characterisation of a bovine cDNA encoding eukaryotic initiation factor 2 alpha

Area of Science:

• Molecular Biology
• Biochemistry
• Genetics

Background:

• Eukaryotic initiation factor 2 alpha (eIF-2 alpha) plays a critical role in protein synthesis regulation.
• Understanding the conservation of eIF-2 alpha across species can provide insights into its fundamental functions.

Purpose of the Study:

• To isolate and characterize cDNA clones encoding the alpha-subunit of bovine eukaryotic initiation factor 2 (eIF-2 alpha).
• To confirm the identity and functionality of the isolated bovine eIF-2 alpha sequence.

Main Methods:

• Isolation of cDNA clones from a bovine lymphosarcoma library.
• Sequence analysis to determine amino acid identity with other species.
• Expression of full-length bovine eIF-2 alpha cDNA in yeast.
• Polypeptide characterization using molecular mass determination and antibody cross-reactivity.

Main Results:

• Two cDNA clones encoding bovine eIF-2 alpha were successfully isolated.
• The predicted amino acid sequence exhibited over 99% identity to rat and human eIF-2 alpha.
• Functional expression in yeast produced a 36 kDa polypeptide that reacted with anti-eIF-2 alpha antibodies.

Conclusions:

• The isolated cDNA clones accurately represent the bovine eIF-2 alpha subunit.
• Bovine eIF-2 alpha is highly conserved among mammals, suggesting a conserved role in translation initiation.
• The study provides a valuable resource for further research into eIF-2 alpha function and regulation in cattle.