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91.
Directed excision of a transgene from the plant genome 总被引:40,自引:0,他引:40
Sandra H. Russell Joyce L. Hoopes Joan T. Odell 《Molecular & general genetics : MGG》1992,234(1):49-59
Summary The effectiveness of loxP-Cre directed excision of a transgene was examined using phenotypic and molecular analyses. Two methods of combining the elements of this system, re-transformation and cross pollination, were found to produce different degrees of excision in the resulting plants. Two linked traits, -glucuronidase (GUS) and a gene encoding sulfonylurea-resistant acetolactate synthase (ALSr), were integrated into the genome of tobacco and Arabidopsis. The ALSr gene, bounded by loxP sites, was used as the selectable marker for transformation. The directed loss of the ALST gene through Cre-mediated excision was demonstrated by the loss of resistance to sulfonylurea herbicides and by Southern blot analysis. The -glucuronidase gene remained active. The excision efficiency varied in F1 progeny of different lox and Cre parents and was correlated with the Cre parent. Many of the lox × Cre F1 progeny were chimeric and some F2 progeny retained resistance to sulfonylureas. Re-transformation of lox/ALS/lox/GUS tobacco plants with cre led to much higher efficiency of excision. Lines of tobacco transformants carrying the GUS gene but producing only sulfonylurea-sensitive progeny were obtained using both approaches for introducing cre. Similarly, Arabidopsis lines with GUS activity but no sulfonylurea resistance were generated using cross pollinations. 相似文献
92.
Joan Sloan Tracy A. Warner Paul T. Scott Trudi L. Bannam David I. Berryman Julian I. Rood 《Plasmid》1992,27(3)
A newClostridium perfringens-Escherichia coli shuttle plasmid has been constructed and its complete DNA sequence compiled. The vector, pJIR418, contains the replication regions from theC. perfringens replicon pIP404 and theE. coli vector pUC18. The multiple cloning site and lacZ gene from pUC18 are also present, which means that X-gal screening can be used to select recombinants inE. coli. Both chloramphenicol and erythromycin resistance can be selected inC. perfringens andE. coli since pJIR418 carries theC. perfringens catP and ermBP genes. Insertional inactivation of either the catP or ermBP genes can also be used to directly screen recombinants in both organisms. The versatility of pJIR418 and its applicability for the cloning of toxin genes fromC. perfringens have been demonstrated by the manipulation of a cloned gene encoding the production of phospholipase C. 相似文献
93.
Pablo Collazo Lluís Montoliu Pere Puigdomènech Joan Rigau 《Plant molecular biology》1992,20(5):857-867
The isolation and characterization of cDNA and homologous genomic clones encoding the lignin O-methyltransferase (OMT) from maize is reported. The cDNA clone has been isolated by differential screening of maize root cDNA library. Southern analysis indicates that a single gene codes for this protein. The genomic sequence contains a single 916 bp intron. The deduced protein sequence from DNA shares significant homology with the recently reported lignin-bispecific caffeic acid/5-hydroxyferulic OMTs from alfalfa and aspen. It also shares homology with OMTs from bovine pineal glands and a purple non-sulfur photosynthetic bacterium. The mRNA of this gene is present at different levels in distinct organs of the plant with the highest accumulation detected in the elongation zone of roots. Bacterial extracts from clones containing the maize OMT cDNA show an activity in methylation of caffeic acid to ferulic acid comparable to that existing in the plant extracts. These results indicate that the described gene encodes the caffeic acid 3-O-methyltransferase (COMT) involved in the lignin biosynthesis of maize. 相似文献
94.
I.-Wu Chen Juinn H. Lin Joan Dorley 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1992,576(2)
A high-performance liquid chromatographic (HPLC) procedure has been developed for the quantification of L-365,260 (I), a cholecystokinin and gastrin receptor antagonist, in dog and rat plasma. The method involves liquid—liquid extraction and HPLC with ultraviolet detection. Standard curves were linear over the range 7.5–2000 ng/ml for rat and dog plasma. The method is reproducible and reliable with a detection limit of 7.5 ng/ml in biological fluids. The mean coefficients of variation for concentrations within the range of the standard curve range were 3.84 and 2.56%, respectively, for intra-day analysis and 4.48 and 4.26%, respectively, for inter-day analysis. Application of the development was successfully demonstrated by quantifying the concentration of I in both dog and rat plasma samples following an intravenous or oral dose of 5 mg/kg I. 相似文献
95.
Scott K. Davis Joan E. Strassmann Colin Hughes L. Susan Pletscher Alan R. Templeton 《Evolution; international journal of organic evolution》1990,44(5):1242-1253
Six variable protein loci and one variable ribosomal DNA restriction site were used for an analysis of population structure in five species of Polistes from Texas. A sample-reuse algorithm was developed that estimated FST, FIS, and ø (the coefficient of kinship) from probabilities of identity. Of the four species analyzed in detail only one, Polistes exclamans, had statistically significant values of FST. These values may reflect natural constraints on successful nesting for migrants of this species. Three of the four species had significant values of FIS and three of the four species had significant values of ø. In many cases ø also differed from the expected value under haplodiploidy and random mating. Values of ø did not differ from expectations under haplodiploidy and local inbreeding. These results emphasize that theories of social behavior and evolution based on coefficients of kinship should include some explicit consideration of population structure. 相似文献
96.
A TLC procedure which resolves two molecular species of ethanolamine plasmalogen, Pl-PE-1 and Pl-PE-2, was used to compare the ratio of these two species in myelin isolated from normal appearing white matter from brains of 17 multiple sclerosis (MS) patients, 17 normal (N) individuals, 1 patient with subacute sclerosing panencepha itis (SSPE) and 1 patient with a non-demyelinating neurological disease (OND). One of these species (Pl-PE-2) has been reported to be unique to myelin and has primarily 181 in both the 1 and 2 positions of glycerol. The other species (Pl-PE-1) is also present in other membranes and has primarily a saturated chain in the 1 position and a polyunsaturated chain in the 2 position. The Pl-PE-1 to Pl-PE-2 ratio was quantitated by scanning the plates with a densitometer. The ratio was similar to normal in most of the MS samples, 0.88±0.09, but was much less than normal in 4 of the MS samples and the SSPE sample. This is attributed to increased decomposition or hydrolysis of Pl-PE-1 relative to Pl-PE-2 at some stage, either during the disease process or due to post-mortem decomposition. Although the reason for the enhanced decomposition of Pl-PE-1 is not known it suggests that Pl-PE-2 is more stable chemically. This may be related to the unique occurrence of Pl-PE-2 in myelin. 相似文献
97.
Implications of a revised United States American Indian nadir population and the pattern of decline leading to it are examined. Substituting the new nadir for that used by Dobyns (1966) lowers by several million his estimates of Indian population for the United States (and Canada) area. These estimates then become more compatible with ones currently being suggested by other scholars. The nineteenth century decline leading to this nadir is found to be remarkably linear. Assuming linearity in decline from initial European contact through the eighteenth century, an aboriginal population estimate of 1,845,183 for the United States area may be extrapolated, along with estimates for intervening years. The nineteenth century decline is then graphed with the data extrapolated to 1492. The resulting pattern of population decline is quite different from the ones suggested by Mooney (1928) and Dobyns (1966). 相似文献
98.
Comparison of the Molecular Forms of the Cholinesterases in Tissues of Normal and Dystrophic Chickens 总被引:7,自引:4,他引:3
Joan M. Lyles Israel Silman Luigi Di Giamberardino† Jean-Yves Couraud † Eric A. Barnard 《Journal of neurochemistry》1982,38(4):1007-1021
Abstract: The levels and molecular forms of acetylcholinesterase (AChE, EC 3.1.1.7) and pseudocholinesterase (ΦChE, EC 3.1.1.8) were examined in various skeletal muscles, cardiac muscles, and neural tissues from normal and dystrophic chickens. The relative amount of the heavy (Hc ) form of AChE in mixed-fibre-type twitch muscles varies in proportion to the percentage of glycolytic fast-twitch fibres. Conversely, muscles with higher levels of oxidative fibres (i.e., slow-tonic, oxidative-glycolytic fast-twitch, or oxidative slow-twitch) have higher proportions of the light (L) form of AChE. The effects of dystrophy on AChE and ΦChE are more severe in muscles richer in glycolytic fast-twitch fibres (e.g., pectoral or posterior latissimus dorsi, PLD); there is no alteration of AChE or ΦChE in a slow-tonic muscle. In the pectoral or PLD muscles from older dystrophic chickens, however, the AChE forms revert to a normal distribution while the ΦChE pattern remains abnormal. Muscle ΦChE is sensitive to collagenase in a similar way as is AChE, thus apparently having a similar tailed structure. Unlike skeletal muscle, cardiac muscle has very high levels of ΦChE, present mainly as the L form; AChE is present mainly as the medium (M) form, with smaller amounts of L and Hc . The latter pattern of AChE forms resembles that seen in several neural tissues examined. No alterations in AChE or ΦChE were found in cardiac or neural tissues from dystrophic chickens. 相似文献
99.
Joan M. Herbers 《Journal of theoretical biology》1981,89(1):175-189
Reliability theory is used to explore the consequences of ant colony's foraging techniques for the evolution of individual competence. Five fundamental strategies are outlined according to Oster & Wilson (1978) and corresponding probabilities of food being returned to the nest are derived. The strategies differ in redundancy schemes and levels of worker reliability. Colony-level selection produces workers of imperfect competence. System reliability can be very high even when the workers are inept, if the foraging strategy is complex and redundant. Individual competence of foragers in a colony depends on diet breadth, caste polymorphism, and tempo. The implications of each variable for colony survival are discussed. 相似文献
100.
Ethanol-induced sleep onset times, sleep times and blood alcohol levels upon awakening were measured in mice fed an essential fatty acid deficient, Purina Chow or unsaturated fat diet for nine months. These values in animals fed the essential fatty acid deficient and Purina Chow diets did not differ, but mice fed the unsaturated fat diet had longer sleep times and lower blood alcohol levels upon awakening than mice fed essential fatty acid deficient or Purina Chow diets. Crude brain mitochondrial fractions isolated from mice fed the essential fatty acid deficient diet had decreased levels of docosahexaenoic [22:6(n-3)] and increased levels of eicosatrienoic [20:3(n-9)], docosatrienoic [22:3(n-9)] and docosapentaenoic [22:5(n-6)] acids compared to mice fed the Purina Chow diet. The unsaturated fat diet decreased 22:6(n-3) and increased 22:5(n-6) compared to the Purina Chow dietary regimen. The longer sleep times and lower blood alcohol levels found in mice fed the unsaturated fat diet probably resulted from an artifact due to the obesity of the mice fed this diet and from the hinderance of obesity to the righting reflex (our measure of ethanol potency). We conclude that the alteration of several polyunsaturated fatty acid components in the brain has little or no influence on the sensitivity of the nervous system to alcohol. 相似文献