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991.
992.
Tihomir Todorov Albena Todorova Bilyana Georgieva Vanyo Mitev 《Molecular biotechnology》2010,45(2):150-154
We report on a unified rapid betaine-based-PCR protocol for amplification of the (CAG)n region in Huntington disease (HD) and the (CGG)n region in Fragile X syndrome (FXS), followed by an electrophoretic separation on automated sequencer for precise determination
of the triplet numbers. The high betaine concentration (2.5 M betaine) permits precise amplification of the CAG and CGG repeats.
Ten HD affected patients and 10 healthy individuals from HD families were re-evaluated. For FXS the CGG region in normal individuals
and premutations of about 100 repeats were precisely amplified by this protocol. Ten unrelated FXS premutation carriers and
24 mentally retarded non-FXS affected boys were re-examined by this method. The results totally coincided with the previous
ones. This protocol is a good choice as a fast screening test. Within 24 h we can have preliminary information on the patient’s
genetic status. Normal individuals, CGG premutation carriers up to 100 repeats, as well as HD patients carrying an expansion
up to 50 CAG repeats can be easily clarified. This accounts for a relatively large proportion (about 90%) of the suspected
HD and FXS patients, referred to our laboratory for genetic analysis. The calculation of the repeat’s number is more accurate
for the correct interpretation of the results, screening tests and genetic counselling. 相似文献
993.
Neopomacentrus filamentosus, a common damselfish on the Indo–Australian archipelago, undergoes significant shifts in size and mitochondrial genetic structure upon larval settlement and metamorphosis to juvenile stages. We characterized five polymorphic microsatellite loci in order to study temporal genetic shifts within a single generation of N. filamentosus sampled first as larval settlers then again as demersal juvenile recruits. All loci were extremely polymorphic and exhibited high levels of heterozygosity. While all loci from the larval samples conformed to Hardy – Weinberg expectations, significant heterozygote deficiencies were seen in two loci in the juvenile samples, likely due to extreme size‐selective mortality imposed post‐settlement. 相似文献
994.
Purification of transcription factor IIIB from HeLa cells 总被引:17,自引:0,他引:17
995.
Metabolism of palmitate in cultured rat Sertoli cells 总被引:1,自引:0,他引:1
Isolated rat Sertoli cells were incubated in the presence of [1-14C]palmitate at a cell concentration of 1.54 +/- 0.31 mg protein/flask (n = 7). The oxidation of palmitate was concentration dependent and maximal oxidation was obtained at 0.35 mM-palmitate. At a saturating concentration of palmitate the oxidation was linear for at least 6 h. About 65% of the total amount of palmitate oxidized during 5 h at 0.52 mM-palmitate (109 +/- 44 nmol/flask, n = 5) was recovered as CO2 and the rest as acid-soluble compounds. Almost all radioactive acid-soluble compounds which were secreted by the Sertoli cells were shown to be 3-hydroxybutyrate and acetoacetate. The palmitate recovery in cellular lipids and triacylglycerols was 9.4 +/- 5.1 nmol/flask (n = 5) and 3.5 +/- 2.8 nmol/flask (n = 5) respectively. Addition of glucose had no significant effect on palmitate oxidation but caused a 9-fold increase in esterification of palmitate into triacylglycerols. We conclude that cultured rat Sertoli cells can oxidize palmitate to CO2 and ketone bodies and that fatty acids appear to be a major energy substrate for these cells. 相似文献
996.
Ni2+ inhibited Ca2+-stimulated adenosine triphosphatase activity in the microsomal fraction of the rat parotid gland in vitro. The Ni2+ concentration required for 50% inhibition was 0.45 mM. Inhibition mechanisms of Ni2+ for Ca2+ and ATP were of the competitive type and the noncompetitive type, respectively. The Ki values of Ni2+ for Ca2+ and ATP were 0.52 and 0.59 mM, respectively. The inhibitory effect of Ni2+ was reversible. 相似文献
997.
2-(2-Pyridyl)ethyl group is a new type P-O protecting group for the synthesis of oligodeoxyribonucleotides by the phosphite triester method. This group is stable to alkali and acid conditions, and to be removed from internucleotidic bonds under mild conditions via two step procedures without any side reactions. Further we have found that bis(diisopropylamino)chlorophosphine is much more effective for the preparation of bis(diisopropylamino)alkoxyphosphines than various dichlorophosphines. 相似文献
998.
N.m.r. studies of metabolism in perfused organs 总被引:1,自引:0,他引:1
J J Ackerman P J Bore D G Gadian T H Grove G K Radda 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》1980,289(1037):425-436
Several metabolites and intracellular pH in intact organs can be studied in a non-destructive manner by phorphorus nuclear magnetic resonance (31P n.m.r.). This possibility was demonstrated by us nearly five years ago. Since then we have developed the appropriate physiological techniques and improved the n.m.r. method for the study of animal hearts and kidneys. Here we described measurements aimed at clarifying three problesm. (1) Having measured the enzyme-catalysed fluxes between phosphocreatine and ATP by the method of saturation transfer n.m.r., we examine the relations between energy supply and heart rate in the isolated perfused rat heart. (2) We describe experiments to establish the validity of the perfusion model. For the first time, we report 31P n.m.r. measurements of an in vivo rat heart and compare the results with those obtained for the perfused rat heart. (3) Ischaemia and metabolism in rabbit kidneys is investigated to establish the relation between functional and metabolic recovery after a renal transplant operation. 相似文献
999.
1000.