R. Jonassen

The ESD polymorphism: Further studies of the ESD2 and ESD5 allele products

SummaryElectrofocusing and agarose electrophoresis techniques both reveal polymorphism of ESD 2, which may be subdivided into two different proteins, coded for by genes allelic to ESD*1. After agarose electrophoresis, ESD 2 is slightly more anodally located than ESD 5, while the latter is considerably more acidic as revealed by electrofocusing in polyacrylamide gel slabs. Family studies have confirmed that each of the allele products behave as Mendelian characters: and the gene frequencies in a Norwegian population material are about 0.08 and 0.02 for the ESD*2 and ESD*5 alleles, respectively.

Restriction fragment length polymorphisms of the complement component C4 loci on chromosome 6: studies with emphasis on the determination of gene number

Restriction fragment length polymorphisms of the C4 region of human chromosome 6 have been studied in family material where the haplotypes are defined with regard to other genetic markers in this region. Employing one near full‐length C4 probe and the combination of BglII and XbaI enzymes, five different C4 genes were characterized. Studies of the segregation of DNA patterns in families made possible the reliable determination of DNA C4 haplotype pattern including gene number. In the total material of 76 haplotypes, 13 different types with regard to number and/or DNA type of C4 gene(s) were encountered. Twelve of the haplotypes had one C4 gene only, 58 had two genes, while 6 had three C4 genes. This fits fairly well with the hypothesis that the one‐ and three‐gene haplotypes have originated through unequal crossing‐over between chromosomes carrying duplicated C4 genes.

GLO Polymorphism in Norway

GLO phenotype distribution and GLO allele frequencies in samples of the Norwegian population and the Lappish minority of Norway are presented. The GLO frequency is 0.442 in 216 Norwegians, while it is 0.304 in 184 Lapps; the difference is statistically significant. There are furthermore probably differences in gene frequencies between two main groups of Lapps.

The human genes for complement components 6 (C6) and 9 (C9) are closely linked on chromosome 5.

We have used a cDNA probe for human complement component 9 (C9), which detects three DNA polymorphisms, to analyse the inheritance of C9 in families informative for C6 protein variants. We found that these genes are closely linked with a lod score of 9.28 at recombination fraction 0.00. There is not indication of allelic association.

The C4 system

SummaryThe present study shows that the C4 system as investigated by high voltage agarose gel electrophoresis is highly polymorphic. In a series of unrelated Norwegian adults, where C4 types have been ascertained through segregation in families, six different haplotypes have been found to occur with a frequency exceeding 1%. The genotype frequencies in the population fit expected Hardy-Weinberg distribution. In family material comprising 89 matings with 327 children the distribution of offspring is as expected according to autosomal codominant inheritance of haplotypes.

Linkage and association studies with C8A and C8B RFLPs on chromosome 1

Linkage relations for the C8A and C8B BamHI RFLPs have been investigated. A peak lod score of 4·52 at recombination fraction zero was obtained between the two C8 genes. Combined with our previously obtained linkage data (Rogde et al. 1986) the maximum lod score is 7·53 at recombination fraction zero. The compiled C8‐PGM1 linkage data from this and the previous study gave a maximum lod score of 22·02 at recombination fraction 0·11 (0·07–0·16) with no sex difference. A chromosome 1p reference marker, D1S57, has been applied in this linkage study. A maximum lod score of 5·06 between the C8 cluster and D1S57 at θ= 0·18 (0·11–0·28) was recorded. The linkage analyses and triply informative families gave evidence that the C8 loci are situated about halfway between PGM1 and D1S57 on the short arm of chromosome 1. There was no evidence of allelic association between the C8A and C8B BamHI RFLPs in 62 unrelated haplotypes.

ESD polymorphism in Norway

SummaryESD phenotype distribution and allele frequencies in 217 Norwegians and 196 Norwegian Lapps are presented. There is good accordance with Hardy-Weinberg distribution, ESD1 allele frequencies are 0.887 in the Norwegians and 0.872 in the Lapps.

Identification in Blood Stains Through DNA Typing with C4 and HLA-DR Probes

A restriction fragment length polymorphism (RFLP) analysis using double digestion of DNA preparations with XbaI and EglII restriction enzymes is presented. In our panel of 46 unrelated individuals 37 different phenotypic patterns were recognized when using C4 and HLA-DR probes. The preliminary discriminative power value when employing both probes (consecutively or simultaneously) is 0.985. In 6 months old blood stains from 7 of the panel members the RFLP patterns were well preserved both in C4 and HLA-DR. The stains from all these individuals were identified when comparing stain DNA patterns with panel control patterns. Based on these laboratory experiments, it is concluded that dNA typing with such probes may become a powerful tool in future stain identification analysis.

Comparison of Cost Benefit between Traditional Paternity Systens and RFLP Analysis. A pilot study

As the basis for a cost/benefit analysis, we have used a material including 20 paternity cases each including mother, child, putative father and two randomly chosen non-fathers.

Fluorescent stains in protein detection on immunoblots

Transfer of proteins to nitrocellulose paper and subsequent staining with an immunoprint technique, is an efficient method for identification of specific proteins as well as for detection of genetic variations. Proteins may be transferred to nitrocellulose paper from different kinds of gels used for separation, and in our experience proteins are readily transferred by passive blotting both from agarose gels and from polyacrylamide gels. The immunoprint technique implies a series of antigen-antibody reactions forming immunocomplexes on the nitrocellulose surface. The first antibody is directed against the protein in question, and protein staining is usually achieved by an enzyme-linked second antibody combined with a suitable substrate. Horseradish peroxidase and alkaline phosphatase are the two most widely used enzymes in commercially available enzyme-linked antibodies. The reaction between enzyme and substrate usually involves the formation of insoluble, coloured products that indicate protein band position on the blot. This staining method is suitable when only one protein on each blot is examined, or when stained proteins are so well separated that their band patterns do not interfere.