Biomedical subjects
Publications and source records attributed to B Brenig.
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Leukemia inhibitory factor receptor (LIFR), epidermal growth factor receptor (EGFR), and their respective ligands have been implicated in regulating growth and development of the early pig conceptus. We isolated a PAC clone containing the porcine gene for LIFR and a BAC clone with the porcine EGFR gene, respectively. On each of these clones one microsatellite marker was identified by sequencing a collection of subclones. These gene-associated markers were evaluated by genotyping of 202 unrelated boars of four different breeds. Based on fluorescence in situ hybridization and radiation hybrid mapping, the porcine LIFR gene was assigned to SSC16q13-->q14. The EGFR gene mapped to SSC9q26.
Twenty paternal half-sib families of a granddaughter design were genotyped for 265 genetic markers, most of them microsatellites. These were 16 Holstein families, 3 Simmental families, and 1 Brown Swiss family. The number of sires per breed was 872, 170, and 32, respectively. Two-point recombination rates were estimated both jointly for all breeds and each single breed separately. Of 1168 marker intervals, 865 provided estimates for at least two breeds. Differences between breeds were tested by likelihood ratio tests. Four marker intervals, representing three genomic regions on BTA19, BTA24, and BTA27, show a significant impact of the breed at a false discovery rate of 0.23 and indicate a genetic component of observed heterogeneity of recombination. The variability of recombination rates between cattle breeds might not be a common feature of the whole genome, but rather might be restricted to certain chromosomal segments. Thus, attention should be paid to heterogeneities when pooling data of such regions from different breeds.
The 3 beta-hydroxysteroid dehydrogenase/Delta 5-Delta 4-isomerase (3 beta-HSD) enzymes are essential for the biosynthesis of steroid hormones. The 3 beta-HSD gene family has been reported to encode for different isoenzymes which function either as dehydrogenase/isomerase or as reductase. The 3 beta-HSD enzymes are involved in the formation of the pheromone androstenone (5 alpha-androst-16-ene-3-one) which contributes to the unpleasant odour present in the meat of uncastrated boars. An reverse-transcription-polymerase chain reaction (RT-PCR) probe from porcine testicular tissue of a 3 beta-HSD enzyme was used to screen a porcine adipose tissue cDNA library. Both strands of the positive clones were sequenced and the putative coding sequence of 1122 nucleotides encodes 374 amino acids. Comparison of the putative open reading frame with the bovine and the human type I homologues revealed 85.6 and 79.3% identity, respectively. Fluorescence in situ hybridization (FISH) was performed with a labelled PAC clone containing the gene of interest. The 3 beta-HSD gene was mapped to the porcine chromosome 4q16-4q21 which is in accordance with the comparative gene map.
Mutations in the gene for gamma-sarcoglycan (SGCG) located on HSA 13q12 are responsible for limb girdle muscular dystrophy (LGMD2C) in human. Here we report the cloning of the canine SGCG gene together with its genomic structure and several intragenic polymorphisms. The coding part of the canine SGCG contains seven exons spanning at least 70 kb of genomic DNA. The chromosome assignment of the canine SGCG gene to CFA 25q21-->q23 confirms that the canine syntenic group 10 corresponds to CFA 25 and also supports the findings of human-canine reciprocal chromosome painting.
The COX7A1 gene encodes a heart- and muscle-specific isoform of the subunit VIIA of cytochrome c oxidase, which is the last component of the mitochondrial electron transfer chain. Cloning and characterization of the porcine COX7A1 gene revealed a highly conserved organization with respect to other mammalian COX7A1 orthologs. The porcine gene consists of four exons spanning approximately 1.5 kb and codes for a peptide of 80 amino acids. The COX7A1 gene showed no variation between pigs from different breeds. The gene was assigned by FISH and RH-mapping to SSC 6q1.1-->q1.2 which is in agreement with previously established comparative maps.
Leukemia inhibitory factor (LIF) is a pleiotropic cytokine involved in early conceptus development in pig. We isolated a PAC clone containing the porcine LIF gene and determined the complete DNA sequence of the gene, which spans about 6.3 kb and consists of five exons including three alternative first exons (1D, 1M, 1T) spliced onto common second and third exons. The LIF-D transcript encodes a protein of 202 amino acids sharing 87, 84, and 78% identity with respectively human, ovine, and murine leukemia inhibitory factors. The LIF-M and LIF-T transcripts both encode a truncated protein of 158 amino acids. Two SNP markers within untranslated regions of the LIF cDNA were identified. One SNP is located in the 5'-UTR of the alternative exon 1T while the other SNP is located in the 3'-UTR of exon 3. Based on fluorescence in situ hybridization and radiation hybrid mapping, the porcine LIF gene was assigned to chromosome 14q2.1-->q2.2.
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Cathepsin L (CTSL) is a lysosomal cysteine protease with potent elastase and collagenase activities. Its high activity in the uterine lumen during the period of placental attachment has led to speculation that CTSL may play an important role during embryonic implantation in the pig. Cathepsins have also been implicated in blastocyst implantation in other species like cat, rat and man. We isolated a PAC clone containing the porcine CTSL gene and determined the complete DNA sequence of the gene, which spans about 5.6 kb and consists of eight exons. The CTSL transcript encodes a primary peptide of 334 amino acids sharing 73-78% identity with other mammalian cathepsin L precursor proteins. Based on fluorescence in situ hybridization and radiation hybrid mapping, the porcine CTSL gene was assigned to chromosome 10q11--> q12.
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We have isolated a recombinant phage harboring the canine CALC-I/alpha-CGRP gene. The gene spans a region of approx. 5.3 kb and consists of six exons with sizes ranging from 95 bp (exon 2) and 494 bp (exon 4). By alternative splicing, two transcripts with ORFs of 390 and 384 nt are generated. These encode either the 32-amino acid-long hormone calcitonin (CALC) or the neurotransmitter calcitonin gene-related peptide (alpha-CGRP) with a length of 37 amino acids after proteolytic processing of precursor molecules. The canine calcitonin precursor consists of 130 amino acids with a molecular mass of 14.05 kDa and a statistical pI of 8.0, whereas the deduced alpha-CGRP precursor harbors 128 amino acids with a molecular mass of 13.87 kDa and a statistical pI of 8.6. Both polypeptides have a common N-terminal region of 76 amino acids that is encoded by exons 2 and 3 and separated by different eight (CALC) or six (alpha-CGRP) amino acid spacers from the biologically active polypeptide. The CALC-I/alpha-CGRP gene is a member of the calcitonin gene family and was assigned to chromosome CFA 16q25.1. A comparative analysis of different dog breeds revealed a breed-specific allelic d(CAGGAG)-hexanucleotide expansion in exon 3. This expansion results in an elongation of the common N-terminal region by two amino acids (glutamine-glutamic acid) and alters the molecular mass to 14.31 kDa (pI 7.9) and 14.13 kDa (pI 8.5) of the calcitonin and alpha-CGRP precursor, respectively.
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