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Published on: November 26, 2018
Cloning and sequence analysis of a cDNA for lymphocyte proliferation potentiating factor of rabbit polymorphonuclear
Abstract:
Complementary DNA for a rabbit PMN-derived lymphocyte proliferation potentiating factor (PMN factor) was cloned from a cDNA library constructed from poly(A)+RNA of early inflammatory exudate PMN. Oligodeoxyribonucleotides as probes for cloning were synthesized according to the previously reported amino acid sequence of the purified PMN factor. This cDNA encodes a 268-residue protein which is homologous to the established structures of human IL 1 beta (74%0 or murine IL 1 beta (71%). The NH2-terminal amino acid sequence of the endogenously produced PMN factor indicates that the mature molecule is made up of carboxy terminal 152 amino acids of the precursor molecule. Taking into consideration the previous studies on biological activities, this cloned PMN factor is therefore considered to be rabbit IL 1 beta.
Insights
Researchers cloned complementary DNA for a rabbit polymorphonuclear (PMN) cell-derived lymphocyte proliferation potentiating factor. This factor is identified as rabbit interleukin-1 beta (IL-1β), homologous to human and murine IL-1β.
Area of Science:
- Immunology
- Molecular Biology
- Biochemistry
Background:
- Polymorphonuclear (PMN) cells produce factors that influence lymphocyte proliferation.
- A specific PMN-derived factor potentiates lymphocyte proliferation, but its molecular identity was unclear.
Purpose of the Study:
- To clone and characterize the complementary DNA (cDNA) encoding the rabbit PMN-derived lymphocyte proliferation potentiating factor.
- To determine if this factor is a rabbit homolog of Interleukin-1 beta (IL-1β).
Main Methods:
- Construction of a cDNA library from early inflammatory exudate rabbit PMN poly(A)+RNA.
- Synthesis of oligodeoxyribonucleotide probes based on the known amino acid sequence of the purified PMN factor.
- cDNA cloning and sequencing to determine the protein's structure and identify homologies.
Main Results:
- A cDNA encoding a 268-amino acid protein was successfully cloned.
- The deduced protein sequence showed significant homology to human IL-1β (74%) and murine IL-1β (71%).
- Analysis of the N-terminal sequence indicated the mature protein comprises the C-terminal 152 amino acids of the precursor.
Conclusions:
- The cloned PMN factor is identified as rabbit Interleukin-1 beta (IL-1β).
- This finding provides molecular evidence for rabbit IL-1β and its role in immune responses involving PMNs and lymphocytes.
