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131.
为获取高质量的大麦幼苗胚芽鞘保卫细胞以进行转录组相关研究,以大麦品种Morex的幼苗胚芽鞘为实验材料,分别对胚芽鞘表皮条撕取方法和保卫细胞分离方法进行优化。结果表明,通过"刮压"技术,不仅可以获得完整的大麦胚芽鞘表皮条,而且缩短了获取的时间;利用纤维素酶R-10与离析酶R-10制备酶解液,同时在酶解液中加入转录抑制剂,并结合表皮条"穿孔"进行辅助酶解,发现酶解温度为30℃、酶解时间为水浴1.5 h时,可以获取大量具有活性的大麦胚芽鞘保卫细胞。 相似文献
132.
自2000年Polejaeva I A获得第1头克隆猪后,短短几年时间全世界已有10多例成功的报道,使得猪的体细胞核移植有了长足的发展,但目前猪的体细胞核移植效率依然低下,人们对核移植中重编程分子机理的认识知之甚少。简要综述了猪体细胞核移植近年来的研究进展,分析了猪核移植中的技术难点和影响因素。 相似文献
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134.
Lee LE Van Es SJ Walsh SK Rainnie DJ Donay N Summerfield R Cawthorn RJ 《Journal of fish diseases》2006,29(8):467-480
Neoparamoeba pemaquidensis is an ubiquitous amphizoic marine protozoan and has been implicated as the causative agent for several diseases in marine organisms, most notably amoebic gill disease (AGD) in Atlantic salmon. Despite several reports on the pathology of AGD, relatively little is known about the protozoan and its relationship to host cells. In this study, an in vitro approach using monolayers of a rainbow trout gill cell line (RTgill-W1, ATCC CRL-2523) was used to rapidly grow large numbers of N. pemaquidensis (ATCC 50172) and investigate cell-pathogen interactions. Established cell lines derived from other tissues of rainbow trout and other fish species were also evaluated for amoeba growth support. The amoebae showed preference and highest yield when grown with RTgill-W1 over nine other tested fish cell lines. Amoeba yields could reach as high as 5 x 10(5) cells mL(-1) within 3 days of growth on the gill cell monolayers. The amoebae caused visible focal lesions in RTgill-W1 monolayers within 24 h of exposure and rapidly proliferated and spread with cytopathic effects destroying the neighbouring pavement-like cells within 48-72 h after initial exposure in media above 700 mOsm kg(-1). Disruption of the integrity of the gill cell monolayers could be noted within 30 min of exposure to the amoeba suspensions by changes in transepithelial resistance (TER) compared with control cell monolayers maintained in the exposure media. This was significantly different by 2 h (P < 0.05) compared with control cells and remained significantly different (P < 0.01) for the remaining 72 h that the TER was monitored. The RTgill-W1 cell line is thus a convenient model for growing N. pemaquidensis and for studying host-pathogen interactions in AGD. 相似文献
135.
In this study, the possible influence of temperature on infectious pancreatic necrosis virus (IPNV)-induced apoptosis in a zebrafish liver epithelium (ZLE) cell line was investigated. At a lower temperature (18 degrees C), there was expression of viral proteins VP2 and VP3 at 4 h post-infection (p.i.). At this time no expression was found in the high temperature group at 28 degrees C. The cell survival ratio was 52 and 18% at 24 and 48 h p.i., respectively, during IPNV infection at 18 degrees C. In addition, we assayed for apoptosis in IPNV-infected cells with terminal deoxynucleotidyl transferase (TdT)-mediated end labelling (TUNEL) of DNA at different dosages of virus. We found a ratio of apoptotic cells of 8 and 25% at 12 and 18 h p.i., respectively, in the multiplicity of infection (MOI) 1 group. The MOI 10 group had 20 and 45% apoptotic cells at 12 and 18 h, respectively. Furthermore, at 18 degrees C IPNV activated the caspase-8 and 3 from 1.5 to 2 times at 12 and 18 h p.i., respectively. Taken together, these findings suggest that successful virus replication occurs at the low temperature (18 degrees C) compared with the non-permissive temperature of 28 degrees C. Thus, IPNV replication is capable of activating caspase-8 and -3 and inducing host apoptosis. 相似文献
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137.
ABSTRACT: In molluscs, mantle epithelial cells secrete organic matrix proteins to form shells. In this study, we established a culture of mantle epithelial cells by using the mantle pallial layer of scallops. We aimed to identify the mantle epithelial cells expressing scallop shell matrix proteins and establish a culture system of epithelial cells. After the mantle pallial layer was carefully isolated from the mantle tissue, explant culture was performed at 4°C. Most cells that migrated from the explant tissue were round cells. Most of the adhered cells retained round morphology, while some of the cells adhered to the dish and showed morphology similar to that of epithelial-like and fibroblast-like cells. When the cultured cells were immunostained with a polyclonal antibody against the shell matrix protein, the antibody recognized many of the adhered cells. An estimation of the number of epithelial cells revealed that approximately 70% of the adhered cells were epithelial cells. This is the first report to describe epithelial cells in cultured mantle cells, which express shell matrix proteins. This culture system may be a useful method for characterization of the mantle epithelial cells. 相似文献
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139.
Jeleel O. Agboola Emma Teuling Peter A. Wierenga Harry Gruppen Johan W. Schrama 《Aquaculture Nutrition》2019,25(4):783-797
The rigid cell walls of microalgae may hinder their utilization in fish feeds. The current experiment assessed the correlation between the accessibility of microalgae nutrients and their in vivo digestibility in African catfish. Nannochloropsis gaditana biomass was subjected to physical or mechanical treatments to weaken its cell wall; untreated—no disruption treatment (UNT), pasteurization (PAS), freezing (FRO), freeze‐drying (FRD), cold pasteurization (L40) and bead milling (BEM). Six experimental diets formulated from differently treated and untreated microalgae (at 30% diet inclusion level) were tested on growth performance and apparent nutrient digestibility (ADCs) in juvenile African catfish. A basal diet (REF) containing no microalgae was used as reference diet. Results showed that biomass gain and feed conversion ratio of fish fed L40 and BEM diets increased by 13% and 11%, respectively, relative to the UNT diet. Additionally, FRD, FRO, L40 and BEM cell wall disruption treatments improved protein digestibility by 0.5%, 5.9%, 8.4% and 16.3%, respectively, compared to the UNT treatment. There was a positive correlation between accessibility of microalgal nutrients and their digestibility in African catfish. Nutrient digestibility of microalgae was dependent on extent of cell disruption. Also, the impact of cell disruption on nutrient digestibility of microalgae differs between African catfish and Nile tilapia. 相似文献
140.
营养盐浓度对棕鞭藻叶绿素荧光参数、细胞密度和总脂含量的影响 总被引:1,自引:0,他引:1
利用生物化学方法和叶绿素荧光动力学分析技术,研究了棕鞭藻在一次性培养过程中,不同营养盐浓度(f/4、f/2、f、2f、4f)对其叶绿素荧光参数、叶绿素含量、细胞密度、单位体积干质量、总脂含量和总脂收获量的影响。试验结果表明,营养盐浓度为2f时,该藻的最终细胞密度、叶绿素含量、单位体积干质量、总脂含量和总脂收获量均显著高于其他各处理组,由此可以看出,在本试验条件下,最适合棕鞭藻生长和油脂积累的营养盐浓度是2f。不同营养盐浓度对棕鞭藻的叶绿素荧光参数有显著影响,培养6~10d,f/4处理组的叶绿素荧光参数(光系统Ⅱ的最大光能转换效率;光系统Ⅱ的潜在活性;光系统Ⅱ的实际光能转化效率;相对电子传递效率和光化学淬灭)最低,其次为f/2处理组(P0.05)。培养7~10d,2f处理组的光系统Ⅱ的实际光能转化效率、相对电子传递效率和光化学淬灭显著高于其他处理组(P0.05)。培养结束时(第10d),各处理组细胞密度和总脂含量依次为:2f4fff/2f/4。相关性分析结果表明,营养盐浓度为f/4~2f时,棕鞭藻的细胞密度、叶绿素含量、单位体积干质量和总脂含量与营养盐浓度均呈显著的正相关;营养盐浓度为2f~4f时,上述各值与营养盐浓度则均呈显著的负相关。 相似文献