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United States Department of Agriculture

Agricultural Research Service

Title: Enzymatic Amplification and Molecular Cloning of Cdna Encoding the Small and Large Subunits of Bovine Interleukin 12

Authors
item Zarlenga, Dante
item Canals, Ana
item Aschenbrenner R,
item Gasbarre, Louis

Submitted to: Biochimica et Biophysica Acta
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 6, 1994
Publication Date: N/A

Interpretive Summary: New and more effective methods for managing and eradicating parasitic infections in animals of veterinary importance are constantly being sought after. Preliminary evidence in the treatment of humans with host proteins (interleukins) involved in modulating immune responses in all larger mammals has provided startling evidence that one such protein (IL-12) is capable of providing a boost to the immune system and assist the host in ridding itself of infections and tumors. To this end, we have cloned the two genes for bovine interleukin 12. Expression of these genes will produce cloned proteins which together can be used as a therapeutic agent for treating cryptosporidiosis and other zoonotic parasitic infections of cattle. This product, if successful, will have a direct impact on increasing food safety and consumer confidence in American beef products.

Technical Abstract: Previous studies, as well as data presented here, strongly suggest that homologous and not heterologous interleukin 12 (IL-12) molecules should be used when studying therapeutic applications of this cytokine. Consequently, cDNA generated from stimulated abomasal lymph node cells was used to amplify, clone and subsequently sequence the 35 kDa and 40 kDa subunits of bovine interleukin 12 (IL-12) using primers derived from semi-conserved regions between human and mouse IL-12 sequences. The deduced amino acid sequence of the 40 kDa subunit demonstrated 84.4% and 67.6% homology with human and mouse sequences respectively, while the deduced sequence of the 35 kDa subunit exhibited comparable similarities to the human 35 kDa subunit (82.2%) but differed significantly (58.6%) from mouse-derived sequences. This data provides an important foundation for studying cytokine profiles and for analyzing the effects of prophylactic applications of this molecule in the attenuation of parasitic infections in cattle.

Last Modified: 4/18/2014
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