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91.
Paul Alan Cox 《Economic botany》1979,33(4):397-399
92.
Isolation and partial purification of the major abundant class rat seminal vesicle poly(A+)-messenger RNA 总被引:1,自引:1,他引:0 下载免费PDF全文
Per-Erik Mansson Donald B. Carter Alan B. Silverberg Douglas B. Tully Stephen E. Harris 《Nucleic acids research》1979,7(6):1553-1565
Total poly(A(+))-RNA (poly(A(+))-RNA(tot)) was isolated from rat seminal vesicle and its size distribution determined by 70% formamide 5-25% sucrose density analysis. One major peak was resolved in the 10-13 S region and accounted for approximately 35% of the total poly(A(+))-RNA applied. Preparative 1% SDS, 5-20% linear sucrose density gradients also resolved a single major peak in the 11S region (poly(A(+))(11S). Analysis of poly(A(+))-RNA(tot) and poly(A(+))-RNA(11S) under denaturing conditions on 2% agarose gel electrophoresis demonstrated two major components in both poly(A(+))-RNA populations. Size estimations for these components are 620 and 540 NT respectively. (3)H-cDNA was made to both poly(A(+))-RNA(tot) and poly(A(+))-RNA(11S). Back-hybridization of poly(A(+))-RNA(tot) and poly(A(+))-RNA(11S) to their respective (3)H-cDNA revealed a highly abundant class representing 41% and 85% of the sequences in their respective (3)H-cDNA's. The highly abundant class corresponded to 3-5 sequences present in 30,000-50,000 copies/cell. Invitro translation of poly(A(+))-RNA(11S) resulted in two major polypeptides coded for by the 620 NT long and 540 NT long poly(A(+))-RNA respectively.Images 相似文献
93.
The incorporation of [35S]methionine into ovalbumin, a protein containing NH2-terminal N-acetylglycine, has been studied in chicken oviduct magnum cells. The purification of [35S]methionine-labeled ovalbumin from total oviduct proteins was accomplished by dialysis of a crude extract at pH 3.6 followed by chromatography on carboxymethyl cellulose. The radioactive ovalbumin eluted from the column in three peaks (P0, P1, and P2-containing 0, 1, and 2 moles of phosphate, respectively, per mole of ovalbumin). The kinetics of labeling of peaks P0 and P1 showed that the ratio of radioactivity in NH2-terminal methionine to total incorporation was greater at 2 min of labeling than at later times. The transient labeling of the NH2-terminus of ovalbumin with methionine indicates that methionine is the initiator amino acid for the synthesis of this protein, which in its mature form contains NH2-terminal N-acetylglycine. 相似文献
94.
95.
Studies with congenic resistant strains of mice indicate that susceptibility to Rous sarcoma virus tumorigenesis is influenced by a gene or genes associated with the major histocompatibility complex (H-2). These genes manifest dominant relative susceptibility. Preliminary studies indicate that the CBA/J strain harbors a gene or genes for relative susceptibility, which are recessive. These results are compared with other studies onH-2-associated genes affecting murine viral oncogenesis. 相似文献
96.
Methane oxidation by washed cell suspensions of Methylosinus trichosporium OB3B was selectively inhibited by 25 compounds, including metal binding components such as carbon monoxide (85% O2: 15% CO), KCN (10-6 M), αα′-dipyridyl (10-4 M), 8-quinolinol (10-4 M), thiosemicarbazide (10-5 M), thiourea (10-5 M), hydroxylamine (10-4 M), histidine (10-2 M), British Anti-Lewisite (5x10-3 M), and miscellaneous known inhibitors of other oxygenases. A role for copper in the methane oxygenase system was suggested by the pattern of inhibition and relief of inhibition by added metal ions. 相似文献
97.
Alan Tin Lun Lam Valerie Ho Svetlan Vassilev Shaul Reuveny Steve Kah Weng Oh 《Cell proliferation》2022,55(8)
ObjectivesInduced pluripotent stem cells (iPSCs) generated by monolayer cultures is plagued by low efficiencies, high levels of manipulation and operator unpredictability. We have developed a platform, reprogramming, expansion, and differentiation on Microcarriers, to solve these challenges.Materials and MethodsFive sources of human somatic cells were reprogrammed, selected, expanded and differentiated in microcarriers suspension cultures.ResultsImprovement of transduction efficiencies up to 2 times was observed. Accelerated reprogramming in microcarrier cultures was 7 days faster than monolayer, providing between 30 and 50‐fold more clones to choose from fibroblasts, peripheral blood mononuclear cells, T cells and CD34+ stem cells. This was observed to be due to an earlier induction of genes (β‐catenin, E‐cadherin and EpCAM) on day 4 versus monolayer cultures which occurred on days 14 or later. Following that, faster induction and earlier stabilization of pluripotency genes occurred during the maturation phase of reprogramming. Integrated expansion without trypsinization and efficient differentiation, without embryoid bodies formation, to the three germ‐layers, cardiomyocytes and haematopoietic stem cells were further demonstrated.ConclusionsOur method can solve the inherent problems of conventional monolayer cultures. It is highly efficient, cell dissociation free, can be operated with lower labor, and allows testing of differentiation efficiency without trypsinization and generation of embryoid bodies. It is also amenable to automation for processing more samples in a small footprint, alleviating many challenges of manual monolayer selection.We have developed an allied protocol for reprogramming, selecting, expanding and differentiating human pluripotent stem cells on Microcarriers (designated as RepMC). This method allows faster reprogramming, selecting 30‐50‐fold more candidates for characterization and also allows us to find high quality candidates that differentiate to cardiomyocytes and blood lineages. Mechanistically, this method appears to accelerate the induction, maturation and stabilization phases of reprogramming. Our findings help simplify the process of deriving and expanding iPSCs for therapeutic applications, offering a robust and scalable suspension platform for large‐scale generation of clinical grade iPSCs. 相似文献
98.
Priya Padmanabhan Mukund R. Shukla J. Alan Sullivan Praveen K. Saxena 《Plant Cell, Tissue and Organ Culture》2017,128(1):145-160
C4 plants can efficiently accumulate CO2 in leaves and thus reduce wasteful oxygen fixation by the RuBisCO enzyme. Three C4 enzymes, namely carbonic anhydrase (CA), phosphoenol pyruvate (PEPC) and pyruvate orthophosphate dikinase (PPDK), were over expressed in Oryza sativa L. ssp. indica var. Khitish under the control of green tissue specific promoters PD54o, PEPC and PPDK, respectively. Integration of these genes was confirmed by Southern hybridization. The relative expression of PEPC, CA and PPDK were, respectively, 6.75, 6.57 and 3.6-fold higher in transgenic plants compared to wild type plants (control). Photosynthetic efficiency of the transgenic plants increased significantly along with a 12?% increase in grain yield compared to wild type plants. Compared to control plants, transgenic plants also showed phenotypic changes such as increased leaf blade size, root biomass, and plant height and anatomical changes such as greater leaf vein number, bundle sheath cells, and bulliform cells. Our findings indicate that the combined over expression of these three enzymes is an efficient strategy for incorporating beneficial physiological and anatomical features that will enable subsequent yield enhancement in C3 rice plants. 相似文献
99.
100.
Shibin Gao Carlos Martinez Debra J. Skinner Alan F. Krivanek Jonathan H. Crouch Yunbi Xu 《Molecular breeding : new strategies in plant improvement》2008,22(3):477-494
Leaf collection from the field, labeling and tracking back to the source plants after genotyping are rate limiting steps in
leaf DNA-based genotyping. In this study, an optimized genotyping method using endosperm DNA sampled from single maize seeds
was developed, which can be used to replace leaf DNA-based genotyping for both genetic studies and breeding applications.
A similar approach is likely to be suitable for all plants with relatively large seeds. Part of the endosperm was excised
from imbibed maize seeds and DNA extracted in 96-tube plates using individuals from eight F2 populations and seven inbreds. The quality of the resultant DNA was functionally comparable to DNA extracted from leaf tissue.
Extraction from 30 mg of endosperm yields 3–10 μg DNA, which is sufficient for analysis of 200–400 agarose-gel PCR-based markers,
with the potential for several million chip-based SNP marker analyses. By comparing endosperm DNA and leaf DNA for individuals
from an F2 population, genotyping errors caused by pericarp contamination and hetero-fertilization were found to average 3.8 and 0.6%,
respectively. Endosperm sampling did not affect germination rates under controlled conditions, although under normal field
conditions the germination rate, seedling establishment, and growth vigor were significantly lower than that of non-sampled
controls for some genotypes. However, careful field management can compensate for these effects. Seed DNA-based genotyping
lowered costs by 24.6% compared to leaf DNA-based genotyping due to reduced field plantings and labor costs. A substantial
advantage of this approach is that it can be used to select desirable genotypes before planting. As such it provides an opportunity
for dramatic improvements in the efficiency and selective gain of breeding systems based on optimum combinations of marker-assisted
selection and phenotypic selection within and between generations. 相似文献