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The lens cytoskeleton
Springer Science and Business Media LLC - - 1981
H. Bloemendal, E.L. Benedetti, Frans C.S. Ramaekers, I. Dunia
The method of single-nucleotide variations detection using capillary electrophoresis and molecular beacons
Springer Science and Business Media LLC - Tập 36 - Trang 1903-1908 - 2008
Jinhui Wang, Wei Wang, Yanhong Liu, Libo Duo, Lijuan Huang, Xiaofeng Jiang
We demonstrate that single-nucleotide variations in a DNA sequence can be detected using capillary electrophoresis (CE) and molecular beacons (MBs). In this method, the region surrounding the site of a nucleotide variation was amplified in a polymerase chain reaction, then hybridize PCR products with each of MBs. The sequences of the PCR products are different at the site of 2,044 in exon of interleukin (IL)-13 which to be identified. Through denaturation, the PCR product became single strand and hybridized with the completely complementary MB. The MB-target duplexes were separated using CE and solution-based fluorescence techniques. The results show that in each reaction a fluorescent response was elicited from the molecular beacon which was perfectly complementary to the amplified DNA, but not from the other MB whose probe sequence mismatched the target sequence. The method of CE based on MBs is able to identify single-nucleotide variations in a DNA sequence and can discriminate the genotyping of the SNP between the homo- and heteroduplexes of DNA fragments.
Conservation of floral, fruit and chromosomal diversity: a review on diploid and polyploid Capsicum annuum complex in India
Springer Science and Business Media LLC - Tập 48 - Trang 5587-5605 - 2021
Timir Baran Jha, Biplab Kumar Bhowmick
Capsicum as a spice crop, has wild and cultivated forms admired globally, including Indian subcontinent with vast climatic ranges. Systematic representation of the Indian Capsicum is required to address species relationships and sustainable agriculture, in face of unpredictable climatic conditions. We have updated the catalogue of Indian ‘C. annuum complex’ with 28 landraces and populations from different agro-climatic regions. The agro-climatic influence on the origin of stable chili landraces in India is remarkable, especially in the North East. The floral and fruit morphotype standards and chromosomal attributes have been considered for four distinct ‘C. annuum complex’ members under three species. The highlights of study are: (1) comparative profiling of Indian Capsicum species revealing less infraspecific variation within C. frutescens and C. chinense than C. annuum, at par with cultivation status, (2) karyotype analysis of some unique diploid landraces of C. annuum, (3) karyotypic confirmation of the polyploid Dalle Khursani landraces exclusive to India. To obtain more information, we attempted to correlate diversity of fruit and floral morphotype with chromosomal diversity. Existence of elite and rare germplasm found in the regional pockets offer great scope for enriching the agricultural tradition. The present dataset may serve as a template to be continuously upgraded by taxonomists, genomicists and breeders.
The type IIS restriction enzyme MmeI can cut across a double-strand break
Springer Science and Business Media LLC - Tập 50 Số 6 - Trang 5495-5499 - 2023
Maliha Tasnim, T. Jacob Selph, Jason Olcott, John E. Hill
Abstract Background

Type-IIS restriction enzymes cut outside their recognition sites, allowing them to remove their binding sites upon digestion. This feature has resulted in their wide application in molecular biology techniques, including seamless cloning methods, enzymatic CRISPR library generation, and others. We studied the ability of the Type-IIS restriction enzyme MmeI, which recognizes an asymmetric sequence TCCRAC and cuts 20 bp downstream, to cut across a double-strand break (DSB).

Methods and results

We used synthetic double-stranded oligos with MmeI recognition sites close to 5′ end and different overhang lengths to measure digestion after different periods of time and at different temperatures. We found that the MmeI binding and cutting sites can be situated on opposite sides of a DSB if the edges of the DNA molecules are held together by transient base-pairing interactions between compatible overhangs.

Conclusion

We found that MmeI can cut across a DSB, and the efficiency of the cutting depends on both overhang length and temperature.

Common polymorphisms (rs2241766 and rs1501299) in the ADIPOQ gene are not associated with hypertension susceptibility among the Chinese
Springer Science and Business Media LLC - Tập 39 - Trang 8771-8775 - 2012
Bo Xi, Dan He, Qijuan Wang, Jian Xue, Man Liu, Jun Li
The ADIPOQ gene has been implicated in the etiology of hypertension. However, the results have been inconsistent. In this study, a meta-analysis was performed to assess the associations of ADIPOQ polymorphisms with hypertension risk among the Chinese. Published literature from PubMed, CNKI and Wanfang Data were retrieved. Pooled odds ratio (OR) with 95 % confidence interval (CI) was calculated using fixed- or random-effects model. Six studies (1,812 cases and 2,631 controls) for rs2241766 polymorphism and four studies (1,449 cases and 2,175 controls) for rs1501299 polymorphism were identified. A marginally significant association was observed for rs2241766 polymorphism under recessive genetic model (GG vs. GT+TT: OR = 1.22, 95 % CI 1.01–1.48) and for rs1501299 polymorphism under heterogeneous co-dominant model (TG vs. GG: OR = 0.86, 95 % CI 0.75–0.99) and dominant model (TT+TG vs. GG: OR = 0.85, 95 % CI 0.74–0.98). In addition, under other genetic models, there was no significant association for rs2241766 polymorphism (GG vs. TT: OR = 1.20, 95 % CI 0.98–1.48; GT vs. TT: OR = 0.97, 95 % CI 0.85–1.10; GG+GT vs. TT: OR = 1.01, 95 % CI 0.90–1.15) and for rs1501299 polymorphism (TT vs. GG: OR = 0.82, 95 % CI 0.62–1.08; TT vs. TG+GG: OR = 0.87, 95 % CI 0.66–1.14). However, the associations above were not robust by sensitivity analysis. The present meta-analysis indicated the limited evidence of the significant associations between ADIPOQ gene polymorphisms and hypertension susceptibility among the Chinese.
messenger RNA-containing ribonucleoprotein from mitochondrial polyribosomes of rat liver
Springer Science and Business Media LLC - Tập 2 - Trang 143-149 - 1975
O. I. Kisselev, B. S. Gaitskhoki, N. A. Klimov
A ribonucleoprotein was released from carefully purified rat liver mitochondrial polyribosomes after dissociation with 1 M potassium chloridepuromycin. This ribonucleoprotein was characterized by a sedimentation coefficient ranging from 10–14 S and buoyant density of 1.48 g cm-3 in cesium chloride equilibirum centrifugation differing in these parameters from the subunits of mitochondrial ribosomes. Poly(A)-containing RNA constituted more than 30% of the total RNA content in this non-ribosomal ribonucleoprotein.
Analysis of the spatial and temporal expression pattern directed by the Populus tomentosa 4-coumarate:CoA ligase Pto4CL2 promoter in transgenic tobacco
Springer Science and Business Media LLC - Tập 40 - Trang 2309-2317 - 2012
Xiang Pan, Huanhuan Li, Hongyi Wei, Wankai Su, Xiangning Jiang, Hai Lu
4-Coumarate:CoA ligase (4CL) is a key enzyme in the phenylpropanoid synthesis pathway. The Pto4CL2 promoter was cloned from Populus tomentosa Carr. and fused to the reporter gene encoding β-glucuronidase (GUS); the complex expression patterns directed by the Pto4CL2 promoter were then characterized in Nicotiana tabacum Xanthi by histochemical assays. The promoter 5′-deletion and histochemical assay conducted on transformants indicated that the −317 to −292 nt region supports Pto4CL2 expression in the epidermis and petals and the deletion of the −266 to −252 nt region resulted in the loss of tissue specificity and a dramatic reduction in GUS activity. Furthermore, electrophoretic mobility shift assays testified that an adenine and cytosine-rich element (−264 to −255 nt) and an abscisic acid-responsive element (−242 to −235 nt) in the Pto4CL2 promoter would have functions for the complex expression profiling and efficient basal expression, respectively. These results further clarify the mode of the regulatory expression of class II 4CL promoters in higher plants.
On the Role of Mi-CK and VDAC in Mitochondrial Function of Heart Muscle Cells
Springer Science and Business Media LLC - Tập 29 - Trang 183-186 - 2002
Frank ter Veld, Frank Bruggeman, Jeroen Jeneson, Klaas Nicolay
A two-compartment kinetic model was used to describe reconstituted systems in which mitochondria compete with pyruvate kinase for kinase-generated ADP. The modelling suggests that cytosolic CK deficiency results in a 50% increase in outer mitochondrial membrane permeability.
Cardiac fibrosis and curcumin: a novel perspective on this natural medicine
Springer Science and Business Media LLC - - 2021
Fatemeh Sadoughi, Jamal Hallajzadeh, Liaosadat Mirsafaei, Zatollah Asemi, Mahdi Zahedi, Mohammad Alì Mansournia, Bahman Yousefi
Differential gene expression in pancreatic tissues of streptozocin-induced diabetic rats and genetically-diabetic mice in response to hypoglycemic dipeptide cyclo (His-Pro) treatment
Springer Science and Business Media LLC - Tập 39 - Trang 8821-8835 - 2012
Song Ah Choi, Hyung Joo Suh, Jong Won Yun, Jang Won Choi
Diabetic studies are mostly interested in gene expression in the pancreas, the site of insulin secretion that regulates blood glucose levels. However, a single gene approach has been ruled out for many years in discovering new genes or the molecular networks involved in the induction process of diabetes. To understand the molecular mechanisms by which cyclo (His-Pro) (CHP) affects amelioration of diabetes mellitus, we performed gene expression profiling in the pancreatic tissues of two diabetic animal models, streptozocin (STZ)-induced diabetic rats (T1DM) and genetically-diabetic (C57BL/6J ob/ob) mice (T2DM). To understand the healing process of these diabetic rodents, we examined the effects of CHP on various gene expression in pancreatic tissues of both animal models. Our microarray analysis revealed that a total of 1,175 genes were down-regulated and 629 genes were up-regulated in response to STZ treatment, and the altered expression levels of numerous genes were restored to normal state upon CHP treatment. In particular, 476 genes showed significantly altered gene expression upon CHP treatment. In a functional classification, 7,198 genes were counted as differentially expressed in pancreatic tissues of STZ- and CHP-treated rats compared with control, whereas 1,534 genes were restored to normal states by CHP treatment. Microarray data demonstrated for the first time that overexpression of the genes encoding IL-1 receptor, lipid metabolic enzymes (e.g. Mte1, Ptdss1, and Sult2a1), myo-inositol oxygenase, glucagon, and somatostatin as well as down-regulation of olfactory receptor 984 and mitochondrial ribosomal protein, which are highly linked to T1DM etiology. In genetically-diabetic mice, 4,384 genes were altered in gene expression by more than 2-fold compared to the control mice, when counted differentially expressed. In genetically-diabetic mice, 4,384 genes altered in expression by higher than 2-fold were counted as differentially expressed genes in pancreatic tissues of CHP-treated mice. On the other hand, 2,140 genes were up-regulated and 2,244 genes were down-regulated by CHP treatment. The results of the microarray analysis revealed that up-regulation of IL-2, IL12a, and leptin receptor and down-regulation of PIK3 played important physiological roles in the onset of T2DM. In conclusion, we hypothesize that CHP accelerates alterations of gene expression in ameliorating diabetes and antagonizes those that induces the disease.
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