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Human L-type Ca2+ channel alpha 1C subunit gene (CACNL1A1) maps to the distal region of chromosome 12p13, and is composed of approximately 50 exons spanning over 150 kb of the human genome as estimated by restriction map analysis. However, the structure and the total length of the 3'-end of the gene is not clear because the size of several big introns remains unknown. Here the fiber-FISH technique was used to determine the relative order and size of eight partial genomic DNA clones from the central and 3'-terminal regions of CACNL1A1. The total physical distance of this region, including the size and gap distances between the clones were re-estimated. The results show that the physical order of the tested clones was 5'-g14-5 > g12-2 > g10-8 > g4-5 > g16-7 > g8-3 > g12-5 > g6-20-3'. Their individual sizes vary between 6.7 and 21.9 kb. Clones g6-20 and g12-5, both containing repetitive exon 45/46-like element, were found to be located within 59.1 kb downstream of g8-3 containing earlier identified polyadenylation site, i.e. 229.5 kb away from clone g14-5 (exons 10, 11). The possible implications of this structural complexity is discussed.

Fluorescence in situ hybridization analysis was carried out on metaphase preparations of a variety of domestic animal species, viz. pigs, cattle, sheep, river buffaloes, swamp buffaloes, horses, and reindeers, using a PCR generated human telomere repeat probe (TTAGGG)n. Three protocols with different hybridization/washing stringencies were applied. Distinct double spots representing the telomeric sites were observed on either ends of the chromosomes in all the species studied, confirming that one-armed chromosomes are not completely telocentric. In pigs, an interstitial telomeric signal was observed on the 6q22 band of all the individuals examined. Although a random variation in the intensity of signals was observed, it was interesting to note that in one of the five cattle studied, very strong hybridization signals were seen on at least three pairs of chromosomes. In sheep, river buffaloes, and swamp buffaloes, where the biarmed chromosomes are considered to be the result of the fusion of 2-3 one-armed chromosomes of the cattle karyotype, no interstitial telomeric signals were observed.

In situ hybridization technique was applied using a tritiated (3H) bovine IFNG cDNA probe to regionally localize the gene on river and swamp buffalo chromosomes. The hybridization signals peaked on the 4q23-->q26 bands in river buffaloes and on the 1p24-->p26 bands in swamp buffaloes. The results are compared to the localization of the same gene in cattle. Possible evolutionary conservation in the Bovidae is discussed.

A approximately 700 bp bovine IFNG cDNA probe was used for the chromosomal localization of the IFNG gene in cattle using in situ hybridization technique. This locus has previously been assigned to chromosome 5 by analysis of a somatic cell hybrid panel. The in situ hybridization results obtained in the present study allow us to map the locus at the 5q22-q24 bands. Relative location of the IFNG gene and the class II cytokeratin and HOX3 gene clusters in cattle and humans is discussed.

Ceruloplasmin (CP) is a copper-binding protein in vertebrate plasma. In the present study, the porcine ceruloplasmin gene was localized to the 13q32-q33 bands by in situ hybridization, using a human CP cDNA probe. This confirmed the localization of the porcine linkage group V to chromosome 13. The CP locus is closely linked to the transferrin locus in pigs. Their relative physical order on chromosome 13 is discussed. Comparisons are made with the order of these two loci in other mammalian species.

Citrate synthase (CS) is a key enzyme of the Krebs tricarboxylic acid cycle. A 1.4 kb porcine CS cDNA probe was used to chromosomally localize the CS gene in pigs by in situ hybridization. Two in situ hybridization experiments were conducted. Although the first experiment indicated a distinct signal on the 5p12-p13 bands, a secondary signal was observed on the 13q24-q32 bands. Hence, a second in situ hybridization experiment was conducted at higher stringency. The results demonstrated a consistent signal on the 5p12-p13 bands, and the signal on chromosome 13 was scattered with no prominent secondary peak. The CS gene was therefore assigned to the p12-p13 bands of chromosome 5 in pigs.

By the use of in situ hybridization, the bovine immunoglobulin gamma heavy chain (IGHG) gene was assigned to chromosome 21q24. Fifteen percent of the total grains were scored on chromosome 21, with about 65 percent of these grains located on the q24 band. The present results confirm a previous synteny study and provide the precise chromosomal localization of this gene in the cattle genome.

Using rat hormone sensitive lipase (LIPE) and human insulin receptor (INSR) cDNA probes, the LIPE gene was assigned to chromosome 6p11-q21 and the INSR gene to chromosome 2q11-q21 in pigs by in situ hybridization. In humans, these two genes are located on the q and p arms of chromosome 19, respectively. The present results provide the first in situ hybridization mapping data for porcine chromosome 2.

A porcine genomic glucose phosphate isomerase (GPI) DNA probe was used for in situ hybridization with metaphase chromosomes in cattle, sheep and goat. The probe gave distinct signals on the q22----proximal part of the q24 segment of chromosome 18, 14 and 18 in cattle, sheep and goat, respectively, indicating the location of GPI gene. The three species belong to the family Bovidae and have high resemblance in chromosome banding patterns. The localization of the GPI locus to the same site on chromosomes with almost similar banding patterns suggests high degree of homology at these sites in the three species. Correlation between banding homologies and possible similarities at the molecular level is discussed.

The glucose phosphate isomerase (GPI) locus is closely linked to the halothane sensitivity locus in pig. The chromosomal localization of GPI in pig was confirmed to 6p12----q21 by using in situ hybridization. Of the total grains, 25 percent were located on chromosome 6, with about 65 percent clustered in the cent----q21 segment, suggesting the presence of the GPI gene on the very proximal part of the q arm. The efficiency of hybridization was found to be affected by ultraviolet irradiation of metaphases for RBA-banding before hybridization. The irradiated metaphases had higher background grains than the non-irradiated metaphases.