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Domest Anim Endocrinol. 2009 Aug;37(2):104-11. doi: 10.1016/j.domaniend.2009.04.001. Epub 2009 May 14.

A miniature condition in Brahman cattle is associated with a single nucleotide mutation within the growth hormone gene.

Author information

1
University of Missouri, Columbia, MO 65211, USA.

Abstract

Miniature Brahman cattle at the USDA ARS Subtropical Agriculture Research Station in Brooksville, FL have normal proportioned growth but are approximately 70% of mature height and weight when compared with Brahman cattle in the same herd. Pedigree analyses suggest that the condition is inherited through a recessive allele. The miniature Brahman cattle in the Brooksville herd have been used for studies of growth and reproduction, but the underlying causative mutation is unknown. Presumably, the miniature condition could arise from a mutation in the GH gene. The objective, therefore, was to clone the GH cDNA from Brooksville miniature Brahman cattle, compare its sequence to normal Brahman cattle, and test the biological activity of the native GH protein. Messenger RNA was isolated from the pituitary, and a cDNA for the protein coding region of the GH gene was amplified by reverse-transcription polymerase chain reaction (PCR) from each of 2 miniature Brahman bulls. The cDNA were cloned into plasmid vectors, and top and bottom strands were sequenced by automated DNA sequencing. The sequence of both cDNA clones derived from miniature cattle differed from Bos indicus GH (GenBank AF034386) at base number 641 because there was a cytosine (C) instead of a thymine (T). The C to T change encoded a mutation (threonine to methionine) at amino acid 200 (T200M mutation). The mutation was confirmed by sequencing of an additional 2 miniature cattle and comparing their sequence to 2 normal cattle. The threonine is located in the fourth alpha helix of GH and is 1 of 8 amino acids that participate in binding of GH to the GH receptor. Twelve miniature Brahman and 9 normal Brahman cattle were tested by using a restriction fragment length polymorphism analysis that employed the BsmBI restriction enzyme (specific for the mutated nucleotide). The 12 miniature Brahman cattle were homozygous for the mutation (-/-). Seven of the normal Brahman cattle were homozygous for the wild-type allele (+/+), and 2 were heterozygous (+/-). When tested in a cell-based bioassay, GH isolated from the pituitary of -/- cattle (n=4) had approximately 60% activity when compared with GH isolated from the pituitary of +/+ cattle (n=5). In summary, miniature Brahman cattle were homozygous for a single nucleotide polymorphism that encodes a mutation in an amino acid involved in binding of GH to the GH receptor. Cattle of normal size had at least 1 copy of the normal GH allele. Threonine 200 in bovine GH is required for normal growth in cattle.

PMID:
19524387
DOI:
10.1016/j.domaniend.2009.04.001
[Indexed for MEDLINE]

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