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Investigation of PGM1(3), PGM1(6), and PGM1(7) variants by isoelectric focusing. Evidence for new subtypes of the PGM1(3) and PGM1(7) alleles.

A total of 345 haemolysates previously phenotyped by starch gel electrophoresis and known to contain the products of the PGM1(3), PGM1(6), and PGM1(7) alleles have been analyzed by thin layer polyacrylamide gel isoelectric focussing in the pH range 5-7. Two common subtypes, 3+ and 3-, of the PGM1(3) allele have been found in a number of Pacific populations. A single form of the PGM1(7) allele was observed in the Western Caroline Islands. In contrast, one of two Indian PGM1(7) variants focussed to a different position when compared with the form found at polymorphic frequency in the Western Caroline Islands. Only one type of the PGM1(6) allele was detected during the present investigation.

Alleles↗

Substrate affinity in PGM1, PGM2, and PGM2 isozymes.

An easy method for routine detection of PGM1, PGM2, and PGM3 isozymes is given. Differences in substrate affinity are discussed. Gene products pgm1 can be differentiated from gene products pgm3 by cofactor requirement.

Electrophoresis, Agar Gel↗

Localization of the human GLO gene locus.

Data on the linkage relation between the GLO locus and the HLA, Bf, and PGM3 loci are presented. The family material includes 49 GLO/HLA-B (and/or Bf) segregating matings with 134 children informative on 199 parental meioses. Of phase-known meioses, 3 are recombinants and 75 nonrecombinants; linkage is therefore proven. From the total material a distance of 2.5 cM between GLO and HLA-B/Bf is calculated; and from the segregation in some informative family groups it is shown that GLO is situated between PGM3 and HLA-B/Bf.

Chromosome Mapping↗

Possible linkage of HL-A and GLO.

In 21 informative families with 60 children, a possible linkage between HL-A and GLO was found (recombination fraction approximatively 0.15). The sequence of the loci on chromosome 6 might be GLO, HL-A, PGM3, MNSs.

Adult↗

The likely region of overlap (LRO) method for physical assignment of loci.

Numerical analysis is applied to physical assignments of loci, providing point estimates, an LRO confidence interval, and a chi(2) test of consistency whether there is a smallest region of overlap (SRO) or not. Results are given for two examples and summarized for 81 loci in man.

Chromosome Mapping↗

Cloning of a gene involved in cellulose biosynthesis in Acetobacter xylinum: complementation of cellulose-negative mutants by the UDPG pyrophosphorylase structural gene.

Three cellulose-negative (Cel-) mutants of Acetobacter xylinum strain ATCC 23768 were complemented by a cloned 2.8 kb DNA fragment from the wild type. Biochemical analysis of the mutants showed that they were deficient in the enzyme uridine 5'-diphosphoglucose (UDPG) pyrophosphorylase. The analysis also showed that the mutants could synthesize beta(1-4)-glucan in vitro from UDPG, but not in vivo from glucose. This result was expected, since UDPG is known to be the precursor for cellulose synthesis in A. xylinum. In order to analyze the function of the cloned gene in more detail, its biological activity in Escherichia coli was studied. These experiments showed that the cloned fragment could be used to complement an E. coli mutant deficient in the structural gene for UDPG pyrophosphorylase. It is therefore clear that the cloned fragment must contain this gene from A. xylinum. This is to our knowledge the first example of the cloning of a gene with a known function in cellulose biosynthesis from any organism, and we suggest the gene be designated celA.

Cellulose↗

Mapping of the linkage group GLO--Bf--HLA-B,C,A--PGM3. 1. Recombination frequencies.

This study confirms close linkage for the GLO--Bf--HLA-B,C,A complex, and proves linkage between the MHC loci and PGM3. For GLO--PGM3 and Bf--PGM3, respectively, loose linkage seems to be likely, and close linkage can be excluded. Our mapping data on chromosome 6 favor the hypothesis that the PGM3 locus is situated on the HLA-A side of the MHC complex. Yet fine-structure mapping should be confirmed only by segregation analyses in crossover families by testing simultaneously all of the relevant marker loci within uniform family material.

Chromosome Mapping↗