Transgenic vertebrates. Conclusions and outlook.
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Animals↗
Biomedical subjects
G Stranzinger
Publications and source records attributed to G Stranzinger.
Mapping of bovine cytokeratin sequences to four different sites on three chromosomes.
The chromosomal location of bovine class I and class II cytokeratin sequences was determined using in situ hybridization and Southern blot hybridization to DNA from hybrid somatic cells. The main signals were found over chromosome region 19q16----qter after in situ hybridization with two probes for the class I cytokeratin gene subfamily (KRT10 and KRT19) and over region 5q14----q23 after hybridization with probes for the class II gene subfamily (KRT1, KRT5, and KRT8). These regions most likely contain the loci of functional cytokeratin genes, with KRT10 and KRT19 mapping to 19q21 and KRT1, KRT5, and KRT8 to 5q21. The in situ hybridization data were corroborated by analysis of a somatic hybrid cell panel. The genes for the class I keratins segregated concordantly with each other and syntenic group U21 but were discordant with the class II keratin genes. The class II keratin genes segregated concordantly with each other and syntenic group U3. Two class II gene probes gave an additional minor signal above chromosome region 5q25----q33 after in situ hybridization, while another class II probe yielded a minor signal above chromosome region 10q31----qter. When the latter probe and an additional linked probe were hybridized to DNAs from a hybrid panel, two independently segregating loci were recognized, one of which cosegregated with the class II subfamily in syntenic group U3 and the other with syntenic group U5. These data confirm the chromosomal assignment of two syntenic groups and allow the assignment of a formerly unassigned syntenic group.
Animals↗
The bovine genome contains polymorphic microsatellites.
Dinucleotide repeats constitute so-called microsatellites of the human and other eukaryotic genomes. Microsatellite polymorphisms can be identified through the amplification of the microsatellite DNA using the polymerase chain reaction (PCR), followed by resolution of the amplified DNA fragments on a polyacrylamide sequencing gel. We performed a preliminary sequence database search to identify bovine sequences containing (CA)n, (AC)n, (GT)n, or (TG)n blocks, with n greater than or equal to 6. This search yielded 10 sequences containing one or two of the specified repeat blocks and often additional dinucleotide repeat blocks. One of the microsatellite-containing regions has been sequenced twice from independent clones and the reported sequences showed variation in the number of repeats. PCR-amplified fragments of another sequence, the gene for steroid 21-hydroxylase, ranged from 186 to 216 nucleotides in 43 unrelated animals. The database search, as well as the hypervariable microsatellite in the bovine steroid 21-hydroxylase gene, indicates that dinucleotide blocks may be an abundant source of DNA polymorphism in cattle.
Animals↗
Rabbit beta-casein promoter directs secretion of human interleukin-2 into the milk of transgenic rabbits.
To test the potential usefulness of transgenic rabbits as production systems for human proteins of pharmaceutical value, we cloned the rabbit beta-casein promoter and fused it to the genomic sequence of the human interleukin-2 (hIL2) gene. Four transgenic female rabbits were tested for expression and biological activity of the foreign protein in their milk. The milk of all four females proved to contain biologically active hIL2. The results show that transgenic rabbits may represent a convenient and economic system for the rapid production of biologically active protein in milk.
Animals↗
The thyroglobulin gene is syntenic with the MYC and MOS protooncogenes and carbonic anhydrase II and maps to chromosome 14 in cattle.
Using a panel of bovine x Chinese hamster hybrid somatic cells, sequences homologous to genes spanning human chromosome arm 8q have been syntenically assigned in cattle. Thyroglobulin (TG), carbonic anhydrase II (CA2), and the protooncogenes MYC and MOS were assigned to a newly identified bovine syntenic group, U23. Additionally, in situ hybridization of the thyroglobulin probe to bovine metaphase chromosomes revealed this syntenic group to be on bovine chromosome 14 and the bovine thyroglobulin gene to reside at 14q12----q15.
Animals↗
Chromosomal localization of the major histocompatibility complex of the horse (ELA) by in situ hybridization.
The first gene assignment to a horse chromosome is reported for equine leucocyte antigen (ELA), the major histocompatibility complex of the horse. A cloned DNA sequence derived from a class I gene of the porcine major histocompatibility complex was used as a probe for an in situ hybridization experiment. We present the regional localization of ELA, using this sequence, to equine chromosome 20q14-q22.
Animals↗
The loci for parathyroid hormone and beta-globin are closely linked and map to chromosome 15 in cattle.
We report linkage of the loci for beta-globin (HBB) and parathyroid hormone (PTH) in cattle and the assignment of both loci to the bovine chromosome region 15q13-q23. Linkage was analyzed in a family of paternal half-sibs by the use of restriction fragment length polymorphisms detected with bovine probes derived from the HBB and PTH genes. The HBB polymorphism was detected by digestion with restriction endonuclease HindIII and the PTH polymorphism with MspI. The maximum lod score for linkage of PTH with HBB was zeta = 4.52 at theta = 0, suggesting very close linkage of the two loci. The finding of the PTH/HBB linkage is corroborated by the physical assignment of both loci to the region 15q13-q23 by in situ hybridization with bovine genomic probes derived from PTH and HBB, respectively. Since HBB and PTH are syntenic in man and mouse, these results in cattle represent another example of conservation of synteny in the evolution of mammalian chromosomes.
Alleles↗
Chromosome configurations and time sequence of the first meiotic division in bovine oocytes matured in vitro.
Bovine oocytes aspirated from small follicles were cultured for various time periods. Subsequently, the oocytes were fixed and stained with Giemsa and analyzed for their chromosome configuration. The appearance of the various chromosome configurations and their time sequence were documented. The influence of the presence or absence of follicle-stimulating hormone (FSH) during culture was studied. No difference was found in the proportion of oocytes completing meiotic maturation in the presence or absence of FSH. On average, 75% of the oocytes reached metaphase II after 20 h. FSH showed two main effects: 1) all the cumulus oocyte complexes incubated longer than 10 h in the presence of FSH showed cumulus expansion, and 2) the time period required for chromosome condensation was prolonged for 3 h in the presence of FSH. However, the time sequence in vitro in the presence as well as in the absence of FSH paralleled the time sequence found in vivo, where variations of several hours have been reported. The delay in chromosome condensation in the presence of FSH was assumed to be due to a transient increase in cyclic adenosine 3',5'-monophosphate in the cumulus oocyte complexes. As demonstrated for FSH, the described culture system allowed the study of individual factors for their influence on meiotic maturation of bovine oocytes.
Animals↗
Porcine malignant hyperthermia carrier detection and chromosomal assignment using a linked probe.
In pigs, the gene for glucosephosphate isomerase (GPI) is linked to the halothane (HAL) gene which is responsible for malignant hyperthermia (MH). A single copy DNA probe, designated GPI8R, has been isolated from a pig genomic library using a porcine GPI cDNA probe. This probe detects, as was the case for the cDNA probe, a five allele polymorphism in SacI and PvuII digested pig DNA. Family studies show that this polymorphism is linked to the HAL locus and hence can be used in carrier detection. In situ hybridization with GPI8R assigned the GPI locus to bands p12-q22 of chromosome 6. We conclude that the HAL linkage group resides on chromosome 6.
Animals↗
Recombination rates and gene order for some serum alpha-protease inhibitors and immunoglobulin heavy-chain allotypes in pigs.
Linkage analysis between the genes coding for immunoglobulin heavy-chain allotypes and variants of some serum alpha-protease inhibitors produced lod scores above the significance limit of 3. The maximum likelihood estimate of the recombination fraction (theta) ranged from 0.15 to 0.20. Since this is the second report on this linkage group in pigs, the linkage is confirmed. Data from appropriate matings are consistent with a gene order of Pi1-Po1A-(Po1B)-Pi2-Igh1.
Animals↗
Tentative chromosomal localization of the bovine major histocompatibility complex by in situ hybridization.
A genetic region, most likely the major histocompatibility complex, was assigned to bands q13-23 of cattle chromosome 23 by in situ hybridization using a cloned DNA sequence of a class I gene of the pig major histocompatibility complex.
Animals↗
Relationships between the H and A-O blood types, phosphohexose isomerase and 6-phosphogluconate dehydrogenase red cell enzyme systems and halothane sensitivity, and economic traits in a superior and an inferior selection line of swiss landrace pigs.
Associations between production traits and the genes for halothane sensitivity (HAL), S, A and H blood group systems and phosphohexose isomerase (PHI) and 6-phosphogluconate dehydrogenase (6-PGD) enzyme systems were investigated in two lines of pigs selected for an index. The phenotypic variance-covariance matrix of the index included backfat thickness and daily gain, whereas the genetic variance-covariance matrix included daily gain, feed conversion and percentage of lean meat. The experiment was conducted at the experimental station of the Institute of Animal Production and has been underway since 1973. The same index was applied but in two opposite directions to give a superior and inferior line in relation to the production traits. One hundred twenty-nine animals of the superior line in the seventh generation and 88 animals of the inferior line in the sixth generation were studied. Forty-two percent (54/129) of the animals of the superior line were halothane-positive. No animals in the inferior line were halothane reactors. Of the halothane-positive pigs, 70.4% (38/54) in the superior line had the HaHa and 94.4% (51/54) had the SsSs genotype, whereas only 4% (3/75) of the HaHa and 12% (9/75) of the SsSs pigs were halothane-negative. By practicing selection at the H and S loci, it seems possible to efficiently reduce halothane sensitivity in Swiss Landrace pigs. In pigs of the superior line, there were significant differences in percentage of lean meat, carcass length, pH1 (pH value at 45 min to 1 h postmortem, M. longissimus) and reflectance values among genotypes of the HAL, S and H systems and among some genotypes of the 6-PGD system. Poorest meat quality, highest percentage of lean meat and shortest carcass length were observed in pigs homozygous for the alleles HALn, Ss, Ha, PHIB and 6-PGDA. In the inferior line, these associations were absent. As the HAL locus is associated with the above mentioned production traits, linkage disequilibria may explain the observed associations among the H, S, PHI and 6-PGD types and production traits.
Alleles↗
[Cytogenetic polymorphism in farm animals and its significance in animal production and veterinary medicine].
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Animals↗
The haematological and cardiac patterns of adaptation of rabbits to 4,500 m simulated altitude and of readaptation to 400 m.
The haemoglobin (Hb), red blood cell count (RBC) and haematocrit (Hct) of 11 male rabbits were measured at weekly intervals during a 5-week exposure period to hypobaric hypoxia (simulated altitude, 4,500 m) and the following 4 weeks after restoring normoxic conditions in a climatic chamber at constant temperature. RBC showed the slowest response, Hct the fastest during adaptation to hypoxia and readaptation to normoxia. The body weight decreased during the 1st week at hypoxia and remained at a reduced level for 2 weeks after the return to normoxia. In female rabbits exposed up to 43 days to hypoxia the heart ventricle quotient (HVQ) (weights of left ventricle + septum/right ventricle) was determined. There was a gradual decrease of HVQ expressing the development of right heart hypertrophy which was not stabilised within the observation period.
Adaptation, Physiological↗
Chromosomal mutations in pigs derived from X-irradiated semen.
Chromosomal mutations (eight translocations, two pericentric inversions, one paracentric inversion) were found in 8 of 30 phenotypically normal pigs derived from semen irradiated at 800 rads. There were no chromosomal mutations detected in pigs derived from semen irradiated at 600 rads. A summary of the spontaneous and induced cases of chromosomal mutations in the family Suidae suggests that chromosomes 1 and 15 are more often involved in rearrangements.
Animals↗
Comparison of chromosomal protein patterns of thymus and liver in cattle, sheep, goats and pigs.
Chromosomal proteins in the thymus and liver tissues of cattle, sheep, goats and pigs were analyzed electrophoretically. Only minor differences in histone H1 subfractions could be detected between the four species. In the 300 mM NaCl soluble fraction of nonhistone chromosomal proteins (NHCP), tissue- and species-specific qualitative and quantitative differences were found in the protein patterns. The potential practical application of the study of chromosomal proteins to animal breeding is discussed.
Animals↗
X-chromosome gene assignment of swine and cattle.
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Animals↗
Mithramycin and DIPI: a pair of fluorochromes specific for GC-and AT-rich DNA respectively.
The AT specificity of the fluorochromes DIPI and DAPI and the GC specificity of mithramycin are evidenced by observations in human, mouse, and bovine chromosomes. DIPI and DAPI produce a pattern similar to Hoechst 33258 in all three species, whereas mithramycin results in a reverse pattern. The AT-rich centromeric heterochromatin in mouse is brilliantly stained by DIPI or DAPI and remains nearly invisible after mithramycin staining. In the GC-rich centromeric heterochromatin of cattle the opposite behavior is observed.
Animals↗