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991.
Hui Liu Yuejing Zhang Chongxi Liu Baozhu Fang Chuang Li Xuejiao Guan Lianjie Li Xiangjing Wang Wensheng Xiang 《Antonie van Leeuwenhoek》2014,106(6):1207-1214
A novel actinobacterium, designated strain NEAU-ML12T, was isolated from a millipede (Kronopolites svenhedind Verhoeff), which was collected from Fenghuang Mountain in Wuchang, Heilongjiang Province, north China. The strain was characterized using a polyphasic approach. Strain NEAU-ML12T was found to have morphological and chemotaxonomic characteristics typical of the members of the genus Rhodococcus. 16S rRNA gene sequence similarity analysis showed that the strain NEAU-ML12T belongs to the genus Rhodococcus, and was most closely related to Rhodococcus tukisamuensis Mb8T (98.9 %) and Rhodococcus koreensis DNP505T (97.7 %). Phylogenetic analysis based on 16S rRNA gene sequences also demonstrated that strain NEAU-ML12T should be classified in the genus Rhodococcus, forming a distinct clade with R. tukisamuensis Mb8T supported by a 99 % bootstrap value. However, the DNA–DNA relatedness between strain NEAU-ML12T and R. tukisamuensis Mb8T was found to be 41.9 ± 0.7 %. Furthermore, strain NEAU-ML12T could also be differentiated from R. tukisamuensis Mb8T and other closely related strains (R. koreensis DNP505T and Rhodococcus maanshanensis M712T) by morphological and physiological characteristics. Therefore, it is proposed that strain NEAU-ML12T represents a novel species of the genus Rhodococcus, for which the name Rhodococcus kronopolitis sp. nov. is proposed. The type strain is NEAU-ML12T (=CGMCC 4.7145T = DSM 46702T). 相似文献
992.
Minghui Li Huihui Yang Jiue Zhao Lingling Fang Hongjuan Shi Mengru Li Yunlv Sun Xianbo Zhang Dongneng Jiang Linyan Zhou Deshou Wang 《Genetics》2014,197(2):591-599
Studies of gene function in non-model animals have been limited by the approaches available for eliminating gene function. The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR associated) system has recently become a powerful tool for targeted genome editing. Here, we report the use of the CRISPR/Cas9 system to disrupt selected genes, including nanos2, nanos3, dmrt1, and foxl2, with efficiencies as high as 95%. In addition, mutations in dmrt1 and foxl2 induced by CRISPR/Cas9 were efficiently transmitted through the germline to F1. Obvious phenotypes were observed in the G0 generation after mutation of germ cell or somatic cell-specific genes. For example, loss of Nanos2 and Nanos3 in XY and XX fish resulted in germ cell-deficient gonads as demonstrated by GFP labeling and Vasa staining, respectively, while masculinization of somatic cells in both XY and XX gonads was demonstrated by Dmrt1 and Cyp11b2 immunohistochemistry and by up-regulation of serum androgen levels. Our data demonstrate that targeted, heritable gene editing can be achieved in tilapia, providing a convenient and effective approach for generating loss-of-function mutants. Furthermore, our study shows the utility of the CRISPR/Cas9 system for genetic engineering in non-model species like tilapia and potentially in many other teleost species. 相似文献
993.
994.
Collective cell migration is critical for normal development, tissue repair and cancer metastasis. Migration of the posterior lateral line primordium (pLLP) generates the zebrafish sensory organs (neuromasts, NMs). This migration is promoted by the leader cells at the leading edge of the pLLP, which express the G protein-coupled chemokine receptor Cxcr4b and respond to the chemokine Cxcl12a. However, the mechanism by which Cxc112a/Cxcr4b signaling regulates pLLP migration remains unclear. Here we report that signal transduction by the heterotrimeric G protein subunit Gβ1 is essential for proper pLLP migration. Although both Gβ1 and Gβ4 are expressed in the pLLP and NMs, depletion of Gβ1 but not Gβ4 resulted in an arrest of pLLP migration. In embryos deficient for Gβ1, the pLLP cells migrated in an uncoordinated fashion and were unable to extend protrusions at the leading front, phenocopying those in embryos deficient for Cxcl12a or Cxcr4b. A transplantation assay showed that, like Cxcr4b, Gβ1 is required only in the leader cells of the pLLP. Analysis of F-actin dynamics in the pLLP revealed that whereas wild-type leader cells display extensive actin polymerization in the direction of pLLP migration, counterparts defective for Gβ1, Cxcr4b or Cxcl12a do not. Finally, synergy experiments revealed that Gβ1 and Cxcr4b interact genetically in regulating pLLP migration. Collectively, our data indicate that Gβ1 controls migration of the pLLP, likely by acting downstream of the Cxcl12a/Cxcr4b signaling. This study also provides compelling evidence for functional specificity among Gβ isoforms in vivo. 相似文献
995.
鸵鸟(Struthiocamelus)属于平胸总目鸟类,雌雄鸵鸟在性成熟前外部形态相同,很难通过外观和形态来鉴定性别,给早期分群饲养造成了很大的困难。实验利用鸵鸟羽毛提取基因组DNA,之后利用EE0.6和CHD基因中2个引物组合对3对已知性别和9只未知性别鸵鸟的性别基因片段进行特异性扩增。结果显示,这对引物组合在雄性鸵鸟的DNA中未扩增出片段,在雌性鸵鸟DNA中扩增出1条片段,可以对鸵鸟的性别作出准确鉴定,从而解决幼雏期鸵鸟难以从外貌上区分其性别的问题。 相似文献
996.
Polymorphisms in the ovine myostatin gene (MSTN) and their association with growth and carcass traits in New Zealand Romney sheep 总被引:1,自引:0,他引:1
J. G. H. Hickford R. H. Forrest H. Zhou Q. Fang J. Han C. M. Frampton A. L. Horrell 《Animal genetics》2010,41(1):64-72
Myostatin is a regulator of myogenesis and has been implicated in the regulation of adiposity and in controlling the structure and function of tendons. Polymerase Chain Reaction Single-Stranded Conformational Polymorphism (PCR-SSCP) analysis of intron-1 was used to identify five variants (designated A-E) of the myostatin gene ( MSTN ). The effect of this genetic variation on growth and carcass traits was investigated in 517 Romney male lambs from 17 sire-lines, born on a South Island New Zealand farm. General linear mixed effect models revealed that the presence of allele A in a lamb's genotype was associated with decreased leg, loin and total yield of lean meat, whereas the presence of allele B was associated with increased loin yield and proportion loin yield (loin yield divided by total yield expressed as percentage). The effect of the number of allele copies present was investigated, and it was found that the absence of A , or the presence of two copies of B , was associated with increased mean leg yield, loin yield and total yield. Two copies of B were also associated with a decrease in proportion of shoulder yield, whereas two copies of A were associated with a decrease in proportion of loin yield. Associations with allele C were not detected. No associations of MSTN variation with birth weight, weaning weight, pre-weaning growth rate, draft age and hot carcass weight (H-W) were detected. These results suggest that variation in ovine MSTN is associated with meat production, but not birth weight or growth rate in New Zealand Romney sheep. 相似文献
997.
The effects of magnetic stimulation on spinal cord injury-induced migration of white matter astrocytes were studied using an established animal model. Ethidium bromide was injected into the dorsal spinal cord funiculus of adult Sprague-Dawley rats on the left side at T10-11. Animals then received 1.52 Tesla-pulsed magnetic stimulation for 5 min at different frequencies (0-20 Hz) for 14 consecutive days. Selected animals received the non-competitive MEK1/2 inhibitor U0126 (10 microM), prior to stimulation at 10 Hz. Lesion volumes were measured in hematoxylin/eosin-stained sections. Expression of glial fibrillary acidic protein (GFAP), microtubule associated protein-2 (MAP-2) and extra-cellular signal-regulated kinasel/2 (ERK1/2) near the epicenter of injury was examined by Western blotting with quantification using an image analysis system. Lesion volumes decreased and GFAP and p-ERK1/2 expression increased with increasing magnetic stimulation frequency (0-10 Hz). MAP-2 expression was not affected at any frequency. Pretreatment with U0126 reduced GFAP and ERK1/2 expression and increased lesion volumes in response to stimulation at 10 Hz. It is concluded that magnetic stimulation increases the migration of astrocytes to spinal cord lesions. Activation of the ERK1/2 signaling pathway is proposed to mediate astrocyte migration and glial scar formation in response to spinal cord injury. 相似文献
998.
Jin Ming Di Jun Pang Qi Peng Sun Yan Zhang You Qiang Fang Xiao Pen Liu Jian Hua Zhou Xing Xing Ruan Xin Gao 《Molecular biology reports》2010,37(4):1849-1855
CpG-oligonucleotides (CpG-ODNs), mimicking bacterial DNA, have recently been shown to stimulate prostate cancer invasion in
vitro via Toll-like receptor 9 (TLR9). Since cyclooxygenase 2 (COX-2), frequently overexpressed in multiple tumor types including
prostate cancer, is a causal factor for tumor development, invasion and metastasis, an interesting question is raised whether
TLR9 regulates COX-2 expression in prostate cancer cells. To address this question, herein we examined COX-2 expression in
PC-3 cells stimulated with different doses and time courses of CpG-ODNs. The regulatory role of NF-κB in TLR9-mediated COX-2
expression was also investigated. CpG-ODN was found to up-regulate the expression of COX-2 in PC-3 cells in a dose- and time-dependent
manner, but have little impact on COX-1 expression. Moreover, CpG-ODN also promoted nuclear translocation and activation of
NF-κB, which appeared to be required for COX-2 induction by CpG-ODN. Overall, TLR9 up-regulates COX-2 expression in prostate
cancer cells, at least partially through the activation of NF-κB, which may be implicated in tumor invasion and metastasis. 相似文献
999.
1000.
Qingyu Lang Haoxing Zhang Jie Li Fang Xie Yifeng Zhang Bo Wan Long Yu 《Molecular biology reports》2010,37(3):1577-1583
The Aurora kinases play a critical role in mitosis and have been suggested as promising targets for cancer therapy due to
their frequent overexpression in a variety of tumors. Compared with established inhibitors of cell division such as the anti-tubulins,
novel agents target mitotic enzymes and show similar efficacy but with fewer side effects. Several small-molecule inhibitors
of Aurora kinases have been developed as anticancer agents, some of which have progressed to early clinical evaluation. Here
we identified 3-hydroxyflavone as a novel Aurora B inhibitor through high throughput screening. 3-Hydroxyflavone showed potent
inhibition to Aurora B with the IC50 on a nanomolar basis in the enzyme-based kinase activity assay. In the cell-based western blotting analysis, 3-hydroxyflavone
dramatically decreased the phosphorylation level of Histone H3 on the site of serine 10, demonstrating the potent endogenous
Aurora B activity inhibition in cell level. The followed cell image analysis provided the consist result. To make it clear
whether 3-hydroxyflavone inhibited Aurora B by direct binding or not, SPR analysis was carried out to measure the affinity
of interaction between Aurora B protein and 3-hydroxyflavone and the result proved the binding with high affinity. Usually
Aurora activity suppression induced cancer cell proliferation inhibition. Colony formation and cell viability with/without
treatment of 3-hydroxyflavone were measured using CCK-8. The growth suppression under 3-hydroxyflavone present and the growth
recovery after being released gave strong evidence that presence of 3-hydroxyflavone efficiently inhibited the fast growth
of cancer cells. 相似文献