Conjugal transfer of plasmid lambda dv.
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Biomedical subjects
Publications and source records attributed to T Mukai.
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In a marked-inversion-balanced lethal system mutations were accumulated at a minimum pressure of natural selection on 2000 second chromosomes of Drosophila melanogaster that originated from 4 stem chromosomes. Five enzyme loci were tested: alpha-glycerol-3-phosphate dehydrogenase (EC 1.1.1.8), malate dehydrogenase (Mdh, EC 1.1.1.37), alcohol dehydrogenase (EC 1.1.1.1), hexokinase-C (Hex-C tentative name), and alpha-amylase (Amy, EC 3.2.1.1). Three band-morph mutants, one at the Mdh locus, one at the Hex-C locus, and one at the Amy locus, were detected out of 1,658,308 allele replications. In addition, 17 null mutants were found. Accepting that the number of structural genes is the same as that of bands in the salivary gland chromosomes, the total mutation rate per generation for all the structural genes in the second chromosomes is estimated to be 0.008-0.040, which is much smaller than that estimated for viability polygenes (0.12-0.17). Thus, it is speculated that most viability and other fitness polygenes are located in controlling regions outside the structural genes.
The Raleigh, North Carolina, population of Drosophila melanogaster was examined for linkage disequilibrium in 1974, several years after previous analyses in 1968, 1969, and 1970. alphaglycerol-3-phosphate dehydrogenase-1 (alphaGpdh-1), malate dehydrogenase-1 (Mdh-1), alcohol dehydrogenase (Adh), and hexokinase-C (Hex-C, tentative name, F. M. Johnson, unpublished; position determined by the present authors to be 2-74.5) were assayed for 617 second chromosomes, and esterase-C (Est-C) and octanol dehydrogenase (Odh) were assayed for 526 third chromosomes. In addition, two polymorphic inversions in the second chromosomes [In(2L)t and In(2R)NS] were examined, and the following findings were obtained: (1) No linkage disequilibrium between isozyme genes was detected. Significant linkage disequilibria were found only between the polymorphic inversions and isozyme genes [In(2L)t vs. Adh, and In(2R)NS vs. Hex-C]. Significant disequilibrium was not detected between In(2L)t and alphaGpdh-1, which is included in the inversion, but a tendency toward disequilibrium was consistently found from 1968 to 1974. The frequency of two-strand double crossovers within inversion In(2L)t involving a single crossover on each side of alphaGpdh-1 was estimated to be 0.00022. Thus, the consistent but not significant linkage disequilibrium between the two factors can be explained by recombination after the inversion occurred. (2) Previously existing linkage disequilibrium between Adh and In(2R)NS (the distance is about 30 cM, but the effective recombination value is about 1.75%) was found to have disappeared. (3) No higher-order linkage disequilibrium was detected. (4) Linkage disequilibrium between Odh and Est-C (the distance of which was estimated to be 0.0058 +/- 0.002) could not be detected (chi(2) (df=1) = 0.9).-From the above results, it was concluded that linkage disequilibria among isozyme genes are very rare in D. melanogaster, so that the Franklin-Lewontin model (Franklin and Lewontin 1970) is not applicable to these genes. The linkage disequilibria between some isozyme genes and polymorphic inversions may be explained by founder effect.
Utilizing 133-xenon gas and a scintillation camera, we examined the distributional relation between ventillation (V) and perfusion (Q) in patients with pulmonary hypertension (PH) comprising mitral stenosis (MS) and left to right shunt disease (shunt). In the case of MS, Q decreased at the dependent part of lung in proportion to the degree of PH, but preserved normal V and resulted in high V/Q value at this region, which was estimated to cause and physiological dead space effect on the gas exchange function. In the case of shunt disease, even with a high degree of PH, reduction of Q at the dependent lung region was not observed with normal preservation of V, thus obviating any type of the V/Q imbalance. Since the washout of xenon gas by resting tidal volume ventilation was observed to be delayed from the dependent lung region, latent ventilatory impairment in this region due to small airway closure at tidal breathing was also suspected.
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recombinant molecules recovered from transformed E. coli cells consisted of a biologically functional bacterial DNA fragment and, excep
Five hundred and two third chromosomes were extracted from a large cage population of Drosophila melanogaster initiated two months after collection of the progenitors near Raleigh, North Carolina in 1970.---Salivary gland chromosomes of 489 chromosome lines were examined and 54 chromosomes were found to carry inversions. The inversions were classified into three polymorphic types [In (3L)P, In (3R)P, and In (3R)C] and two unique types. The polymorphic inversions were found in frequencies of 0.012, 0.88, and 0.010, respectively.--Viabilities of homozygotes and heterozygotes were examined. Chromosomes with lethals occurred with a frequency of 0.495: 0.537 in the group of inversion-carrying chromosomes and 0.490 in the group of inversion-free chromosomes. The average homozygote viability computed on the basis of an average heterozygote viability of 1.0000 was 0.3235 if lethal lines were included and 0.6290 if they were excluded. The detrimental load to lethal load ratio (D:L ratio) was 0.70 (=0.4636-0.6650). The average viability of lethal heterozygotes was significantly larger than that of lethal-free heterozygotes. It appears, however, that lethal genes in heterozygotes have deleterious effects on fitness as a whole.--The average degree of dominance for viability polygenes was estimated to be about 0.3-0.4 in lethal-free individuals and nearly zero in lethal heterozygotes. Overdominance or some form of balancing selection was suggested at some loci. The difference between the values obtained for average degree of dominance due to genetic backgrounds and superior vibaility of lethal heterozygotes (but not fitness as a whole) suggest that some epistasis or coadaptation occurs.--The results described above are similar to those obtained for the second chromosomes.
A method of in vivo calculation of liver volume using a gamma camera and a 1600 or 4096 channel analyser is described. Topographically, the maximum thickness giving the highest counts in matrix data in the right anterior oblique projection is measured as the maximum breadth in the right posterior oblique projection. Then the total counts given by the liver can be transformed to the volume. The necropsy criteria indicated that the errors were between --11-9 +/- 14-7% in 13 cases by the use of 198Au-colloid. In 20 selected normal control subjects, the mean of measured liver volumes was 779 +/- 99 cm3/m2 of surface area. The volumetric calculation using 99mTc-Sn-colloid or 99mTc-phytate was compared with 198Au-colloid in the same patients. The results with 99mTc-Sn-colloid were well correlated with those with 198Au-colloid, but the results with 99mTc-phytate and 198Au-colloid were almost equal. It was concluded that, excluding rare severe cases of liver cirrhosis, liver volume can be measured with reasonable accuracy.
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In the process of testing whether or not the independent-locus selection model holds true with previously estimated genetic parameters (cf. MUKAI and MARUYAMA 1971) in D. melanogaster collected near Raleigh, North Carolina, we found an abnormal phenomenon: an unusually large increase in dominance variance for viability in comparison with additive variance with the accumulation of mutations on 140 randomyl sampled, inversion-free second chromosomes. Mutations were accumulated only through males heterozygous for the Pm-carrying chromosome [In(2LR)bwV1] and the extracted second chromosomes, and the genetic variance components were estimated by using a partial diallel cross method.--Further investigations clarified that chromosome abberations occurred at a frequency of 0.0114 per second chromosome per generation (inversions: 0.0098; transpositions: 0.0011; translocation: 0.0004), and recessive lethal mutations occurred at an average rate of 0.031 per second chromosome per generation.--From these results and from the amount of change in the homozygous load, it was speculated that about 60--70% of the second chromosomes used had a kind of mutator which induced chromosome and/or chromatid breaks at a minimum rate of 0.18 per second chromosome per generation. These breaks resulted in recessive lethal mutations at a rate more than ten times higher than the normal rate. Also these breaks were most probably the cause of male recombination.--The above unusual increase in dominance variance can be explained by assuming that chromosome segments, introduced into the extracted "wild" chromosomes by male recombinations (double crossover) from the marker chromosomes [In(2LR)bwV1], showed heterosis and linkage disequilibria with deleterious mutations and possibly with other introduced segments.--Finally, the nature and possible significance of mutator factors are discussed.
About 20 copies of plasmid lambda-dv are perpetuated per host chromosome in Escherichia coli K12 cells. The mode of DNA replication of this plasmid lambda-dv was studied, using the density-labelling technique followed by banding in DsCl. It was shown that all the copies of plasmid lambda-dv are potentially capable of replicating roughly once a cell generation. Their replication occurs by random, that is a copy of the plasmid is taken out at random for replication from a pool, to which the two replicas resulting from replication are returned. Chloramphenicol did not inhibit the initiation of a new round of replication of the plasmid molecules, indicating that selection from the replication pool does not require concomitant protein synthesis.
Plasmid lambdadv1, which is in a dimeric form, was converted to a linear monomer duplex by the action of EcoRI restriction endonuclease that incises at a unique site in this plasmid genome. The resulting products were then joined by Escherichia coli DNA ligase to produce molecules with various oligomeric forms, and from these monomeric, dimeric, or trimeric circular molecules were purified. By transformation of cells with these DNAs, clones were obtained that carried lambdadv1 in a monomeric or dimeric form. The former type of clones have not been generated in vivo, except for one in a different host strain, and carriers of timeric or tetrameric lambdadv1's have not been obtained so far. It was observed that a considerable fraction of these oligomeric circular DNAs were converted to lower oligomers (e.g., from trimer to dimer) during transformation. The characteristics of the monomeric lambdadv1 carriers obtained were compared with those of dimeric lambdadv1 carriers. The stabilities of the plasmids of the two forms were the same. However, the monomeric plasmid carriers were less tolerant to lambdavir phage infection and perpetuated about 30% less plasmid genomes in monomer units. Furthermore, dimeric plasmid carriers appeared spontaneously and accumulated in cultures of the monomeric lambdadv1 carriers.
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In order to obtain spatial distribution of an index for regional blood flow at each element on a scintigraphic image of the kidney, we attempted the construction of the so-called functional image. After injecting a single bolus of 133Xe into a renal artery by means of a catheter, this objective was accomplished using digital computer processing for a sequence of scintillation camera recordings of the following washout process from the kidney. This is expressed in a form of matrix of disappearance rate constant. Calculation for the rate constant, the flow index of the functional image, was done using either the least squares (LS) method or height-over-area (H/A) method. Although the former method was considered to be theoretically suitable without undue participation of background activities, the latter was preferred because of stable results for image construction. On reviewing the functional image thus obtained from 22 patients representing a variety of renal diseases, the H/A gave specific information concerning regional distribution of the perfusion integrity mainly related to the cortical part of the kidney. This is often difficult to accomplish utilizing the conventional method of compartmental analysis of the xenon washout curve or selective renal angiography.