Transfer of LM(TK-) cells from normal growth medium to medium lacking K+ leads to a rapid loss of intracellular K+, which is 50-70% inhibited by furosemide or bumetanide. The diuretic-sensitive component of K+ efflux requires both Na+ and Cl-, and is presumably mediated by a K+, Na+, Cl- cotransport system of the kind described in avian erythrocytes and Ehrlich ascites cells. It can be calculated that such a system should be near equilibrium under normal growth conditions but should mediate net efflux (as observed) when the driving force is altered by reducing extracellular K+. The diuretic-sensitive component of net K+ efflux is also sensitive to amiloride. This effect is probably indirect, however, with amiloride acting to block the Na+ influx that supplies Na+ to the cotransport system. At the low extracellular K+ concentrations employed in these studies, the diuretic-sensitive system is a physiologically important pathway of K+ loss. The rate of growth in low-K+ medium can be increased (or the rate of cell lysis decreased) by adding diuretic or by reducing external Na+ or Cl-. 相似文献
The genes governing three of the enzymes of the isoleucine-valine biosynthetic pathway form the operon: operator-ilvA-ilvD-ilvE. The enzymes are: ilvA, l-threonine deaminase; ilvD, dihydroxy acid dehydrase; and ilvE, transaminase B. A nonsense mutation in the ilvD gene (D-ochre) and a nonsense mutation in the ilvE gene (E-amber) affect the properties of the proximal gene product, l-threonine deaminase (TD), in addition to inactivating the enzymes produced by the genes in which the mutations have occurred. The D-ochre mutation causes TD to move in diffusion and gel filtration experiments as though it were 30% smaller than the wild-type enzyme. The E-amber mutation causes TD to move in similar experiments as though it were much larger than the wild-type enzyme. Both mutations completely abolish the sensitivity of TD to l-isoleucine, the normal feedback inhibitor of the wild-type enzyme. The effects of the nonsense mutations on TD can be reversed in three ways: by genetic reversion of the D-ochre mutation; by treatment of the altered enzymes with 3.0 m urea; and by forming a heterozygous diploid, containing the wild-type allele as well as the mutant allele of ilvD or ilvE. The results suggest that the subunits of TD undergo abnormal aggregation in the presence of the partial polypeptides produced by the mutant alleles of ilvD or ilvE; multi-enzyme aggregates in extracts of wild type, however, could not be detected. 相似文献
In Vitro Cellular & Developmental Biology - Plant - Micropropagation is a preferred method to propagate clean, clonal stock plants. Subculture is labor intensive and costly. In vitro hedging... 相似文献
Three streptomycin-suppressible lethal mutants of Escherichia coli K-12 have been shown to possess structurally altered glutamyl-transfer ribonucleic acid (tRNA) synthetases. Each mutant synthetase displays a K(m) value for glutamate which is 10-fold higher than the parental value, and the mutations reside in two widely separate loci on the genetic map. Mixing of the mutant extracts in pairs gave no indication of in vitro complementation. All three enzymes charge the minor tRNA(glu) fraction identically, but one (EM 120) charges the major fraction at a twofold lower rate than do the other two (EM 102 and EM 111). Possible explanations for the existence of the two synthetase loci are presented. 相似文献
Forty-one mutants have been isolated which require streptomycin for growth on complete medium. These streptomycin-suppressible lethal mutations are located randomly around the Escherichia coli genetic map; during growth in liquid culture, they exhibit a variety of responses to the removal of streptomycin as judged by turbidity, cell morphology, and macromolecular synthesis. In particular, some mutants are primarily affected in protein or ribonucleic acid (RNA) synthesis (or both), one in deoxyribonucleic acid synthesis, and two in lipid synthesis. Ten mutants affected in protein synthesis were examined for the activities of all twenty aminoacyl-transfer RNA synthetases, and three were found to have altered glutamyl-transfer RNA synthetase activities. The advantages of this method for isolating a wide variety of conditional lethal mutants are discussed. 相似文献
The aim of this investigation was to establish environmental factors which promote growth and photosynthesis of melon (Cucumis
melo L.) shoot buds, in vitro, and determine if photoautotrophic shoots had superior root forming ability in photoautotrophic
environments. Buds from the triploid melon clone ‘(L-14×B)×L-14’ were observed for 21 days after transfer from a multiplication
MS medium with 3% sucrose and 10 μM benzyladenine (BA) to a shoot development medium with 1 μM BA at three levels of sucrose
in the medium (0, 1 and 3%), and light (50, 100 and 150 PPF) and CO2 (500, 1000 and 1500 ppm) in the culture chamber. More shoot buds were observed with 3% sucrose in the medium. Increased light
and CO2 had a positive interaction with shoot proliferation. Fresh and dry weights were greatest at 3% sucrose, 150 PPF light and
1500 ppm CO2. Shoot buds grew more slowly in sugar-free medium, but fresh and dry weight still doubled over 21 days of culture. Net photosynthetic
rates (NPR) of buds were negative after four days in treatment conditions, but became positive after transfer to fresh, sugar-free
medium. Two triploid genotypes of melon were (1) grown in vitro with sugar (photomixotrophic) and without sugar (photoautotrophic),
(2) rooted in sugar-free media, both in a laboratory controlled environment chamber (in vitro) and a greenhouse acclimatization
unit (ex vitro), and (3) compared for subsequent nursery growth in the greenhouse unit. The genotype ‘(L-14×B)×L-14’ produced
more shoots than ‘(L-14×B)×Mainstream’ in both photomixotrophic or photoautotrophic conditions. ‘(L-14×B)×L-14’ rooted as
well from either photoautotrophic and photomixotrophic shoots but ‘(L-14×B)×Mainstream’ rooted less frequently from photoautotrophic
shoots. Seventy-six percent of the shoots in the laboratory controlled environment chamber were able to root photoautotrophically,
whereas 47% of the shoots in the greenhouse acclimatization unit were rooted. Between 77% and 88% of plantlets from all treatment
combinations survived transfer to the nursery. After growth in the nursery, the sizes of plants (fresh weight, dry weight,
leaf area) were the same for either genotype, from either photoautotrophic or photomixotrophic shoots. Nursery plants that
had been rooted in the laboratory controlled environment chamber were larger than those rooted in the acclimatization greenhouse
chamber.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
LLC-PK1 cells can be brought into a classical quiescent state by depriving them of serum for 6 days. At this time, pulse-labeling with [3H]-thymidine shows that only 3% of the cells are synthesizing DNA, but the quiescent cells can be stimulated with serum to re-enter the cell cycle at a point early in G1. The rate of amiloride-sensitive 22Na+ uptake (as a measure of the Na+/H+ antiporter) is relatively low during quiescence; it rises 2- to 3-fold within 4 h after serum addition. This increase in antiporter activity appears to be required for the resumption of DNA synthesis in the absence of bicarbonate, because ethylisopropylamiloride (EIPA) blocks [3H]-thymidine incorporation when serum is added to cells in bicarbonate-free medium. In the presence of bicarbonate, however, EIPA has no effect on [3H]-thymidine incorporation, indicating that another (bicarbonate-dependent) transport system can substitute for the antiporter under these conditions. 相似文献
Petunia × hybrida Vilm. is a fast-growing ornamental plant that was cultured under varying storage conditions to address seasonal fluctuations in microcutting demand. The effects of storage period (16 to 32 wk), temperature (15 to 23°C), low and high light intensity, sucrose (1 to 5% w/v), and mannitol (0 to 4% w/v) in factorial arrangements were analyzed. Stored and non-stored shoots were compared for microcutting production, harvested twice at 3-wk intervals, and were subsequently transferred to the greenhouse for 17 d. Nearly all plants from 16-wk storage survived well at all treatment conditions and the quality and quantity of microcuttings were enhanced from shoots stored at 15°C and 3% (w/v) sucrose, without mannitol for all storage periods. Another experiment tested 11- to 25-wk storage period with Petunia hybrida ‘Ragtime’ and ‘Suncatcher’ at 12°C and low light intensity. Repeated cycles of microcutting at 2-wk intervals were extended with ex vitro rooting in the greenhouse for 15 d. More and better quality microcuttings were obtained from the second and third cutting cycles than from the first or fourth cycles. By reducing temperature and light intensity, Petunia hybrida was successfully stored for 32 wk (without mannitol). A seasonal schedule with a short production window, followed by cutting large numbers of high-quality shoot-tips, could be affected and efficiently managed through storage. The value of stored shoots was enhanced by extending the number of times a shoot could be cut over repeated cutting cycles with a gain in microcutting quality.