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用带有鸡溶菌酶基因增强子（Ｅ）和启动子（Ｐ）的编码细菌氯霉素乙酰基转移酶（ＣＡＴ）的融合基因（ＥＰＣ），在其上游区、下游区或在上游和下游区同时插入鸡溶菌酶基因３＇端核基质附着区（ＭＡＲ）的不同表达载体ＭＥＰＣ、ＥＰＣＭ和ＭＥＰＣＭ，稳定转染鸡ＨＤ１１Ｐｒｏｍａｃｒｏｐｈａｇｅ同源细胞系，筛选不同表达载体稳定整合后的细胞株，检测ＣＡＴ表达水平，确定其表达基因拷贝数，从而分析鸡溶菌酶基因３＇端ＭＡＲ对基因表达的影响。稳定整合表达和瞬时表达实验结果表明，鸡溶菌酶基因３＇端ＭＡＲ在同源细胞系对基因表达没有激活作用，与该基因５＇端ＭＡＲ在同源或异源细胞系中能激活基因表达形成鲜明对照，说明５＇瑞ＭＡＲ和３＇端ＭＡＲ在调节鸡溶菌酶基因表达上存在一种协调关系。 相似文献
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本研究合成了两个成熟人ＩＬ－６ｃＤＮＡ引物，采用ＰＣＲ法扩增出了成熟人ＩＬ－６基因片段，并在其５＇端加入了一个ＥｃｏＲＩ部位和ＡＴＧ，３＇端加入了一个ＢａｍＨＩ部位和ＴＡＡ。利用ｐＢＶ２２０质粒构建成功了ｐＢＶ２２０ＩＬ－６表达载体。将重组质粒导入Ｅ．ｃｏｌｉＤＨ５ａ中，经３０℃扩增的４２℃诱导，表达出了分子量为２１０００的预期蛋白。经ＳＤＳ－ＰＡＧＥ分析与溥层证明具有明显的ＩＬ－６活性。实验还对 相似文献
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试验以及玉米秸袋模拟地上堆贮进行添加种类筛选和用量研究，试验设立９个处理，分别为按鲜草重加甲酸１．５，３．０，４．５ｇ／ｋｇ，加甲醛１．５，３．０，４．５ｇ／ｋｇ，加甲酸１．５ｇ／ｋｇ＋甲醛１．５ｇ／ｋｇ加甲酸１．５ｇ／ｋｇ＋尿素５．０ｇ／ｋｇ加甲醛１．５ｇ／ｋｇ＋尿素５．０ｇ／ｋｇ。另设不加任何添加剂的对照。结果表明，玉米告青贮甲酸可作为添加剂，按每千克鲜玉米秸添加３ｇ，亦可同时添加尿素５．０ｇ 相似文献
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对两例银黑狐雌性生殖器官形态的观察，卵巢呈两端大小一样的蚕豆形，长１．１－１．３ｃｍ，宽０．７－０．８ｃｍ，厚０．５－０．７ｃｍ。两侧输卵管全长３．０－４．０ｃｍ，有３－１０个弯曲，输卵管有系膜固定。子宫角呈圆筒形，平均长度６．４ｃｍ，外壁直径０．４ｃｍ。子宫体亦为圆筒状，比子宫角粗，长１．４－１．５ｃｍ，直径０．５－０．６ｃｍ，平均体与两子宫角之间形成“丫”状，子宫角内未发现子宫阜。阴道呈扁圆筒 相似文献
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产气荚膜梭菌α毒素保护性抗原基因的克隆与核苷酸序列分析 总被引：19，自引：5，他引：14
将含产气荚膜梭菌α毒素基因的质粒ｐＫＭＡ１００用限制性核酸内切酶ＢａｍＨＩ和ＨｉｎｄⅢ双酶切,经１．５％琼脂糖凝胶电泳分离、透析袋电洗脱法回收０．９５ｋｂα毒素基因片段,再将载体ｐＥＴ－２８ｂ（＋）用ＢａｍＨＩ和ＨｉｎｄⅢ双酶切,然后将处理好的ｐＥＴ－２８ｂ（＋）与０．９５ｋｂ的α毒素基因片段通过Ｔ４ＤＮＡ连接酶进行粘性末端连接,转化至受体菌ＢＬ２１（ＤＥ３）中。经ＢａｍＨＩ和ＨｉｎｄⅢ双酶切分析和核苷酸序列分析,证明重组质粒ｐＸＥＴＡ１含有产气荚膜梭菌α毒素基因。确定了α毒素基因的全部核苷酸序列,并证明其具有正确的阅读框架。 相似文献
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对青海省乐都县５１头驴的１３项血清（浆）生化参数进行测定，结果：总蛋白为７４．４±９．７ｇ／Ｌ，Ａ／Ｇ为Ｏ．９±０．２，总脂为２．５±０．６ｇ／Ｌ，胆固醇为２．５±０．９ｍｍｏｌ／Ｌ，Ｋ￣＋为４．５±０．５ｍｍｏｌ／Ｌ，Ｎａ￣＋为１３７±３ｍｍｏｌ／Ｌ，Ｃａ￣（２＋）为２．３±０．３ｍｍｏｌ／Ｌ，Ｍｇ￣（２＋）为１．６±０．４ｍｍｏｌ／Ｌ，无机磷为２．０±０．６ｍｍｏｌ／Ｌ，ＧＰＴ为８．４±６．４ＩＵ／Ｌ，ＧＯＴ为１１．６±９．７ＩＵ／Ｌ，血浆葡萄糖为５．６±０．５ｍｍｏｌ／Ｌ；上述参数在骟、母驴间无显著差异。 相似文献
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本试验旨在研究饲粮添加不同水平的过瘤胃保护性精氨酸（ｒｕｍｅｎ ｐｒｏｔｅｃｔｅｄａｒｇｉｎｉｎｅ，ＲＰ Ａｒｇ）和大豆油对细毛羊肌内脂肪和脂肪酸组成的影响。试验选用健康、体重相近的 １５只细毛羊，随机分为５组，对照组饲喂基础饲粮，试验组分别饲喂在基础饲粮中添加１．５ｇ／ｄＲＰ Ａｒｇ、２．０ｇ／ｄＲＰ Ａｒｇ、１．５ｇ／ｄＲＰ Ａｒｇ＋３％大豆油和 ２．０ｇ／ｄＲＰ Ａｒｇ＋３％大豆油的饲粮。预试期为５ｄ，正试期为 ４５ｄ。试验结束后每组选 ２只羊进行屠宰，取背最长肌样用于测定肉质指标和脂肪酸组成。结果表明：试验组肌肉 ｐＨ、系水率和熟肉率与对照组差异不显著（Ｐ＞０．０５），而肌内脂肪含量显著高于对照组（Ｐ＜０．０５）；试验组肌肉多不饱和脂肪酸含量与对照组相比显著提高（Ｐ＜０．０５）；添加 ＲＰ Ａｒｇ和大豆油有提高肌肉过氧化物酶体增生物激活受体 γ（ＰＰＡＲγ）、脂蛋白脂酶（ＬＰＬ）ｍＲＮＡ表达量，降低激素敏感性甘油三酯脂肪酶（ＨＳＬ）ｍＲＮＡ表达量的趋势。综合指标，以 １．５ｇ／ｄＲＰ Ａｒｇ组与 １．５ｇ／ｄＲＰ Ａｒｇ＋３％大豆油组添加效果较好。 相似文献
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聂庆华 《江西畜牧兽医杂志》1999,(6):12-13
本试验根据Ｌａｍｂ等［１］报道的鸡生长激素基因序列，设计合成２对特异性引物ＰＭ３和ＰＭＰＳ１，它们的碱基序列分别是ＰＭ３ＦＯＲ５′－ａｔｃｃｃｃａｇｇｃａａａｃａｔｃｃｔｃ－３′ＲＥＶ５′－ｃｃｔｃｇａｃａｔｃｃａｇｃｔｃａｃａｔ－３′ＰＭＰＳ１ＦＯＲ５′－ｃｔａａａｇｇａｃｃｔｇｇａａｇａａｇｇｇ－３′ＲＥＶ５′－ａａｃｔｔｇｔｃｇｔａｇｇｔｇｇｇｔｃｔｇ－３′。通过ＰＣＲ反应从鸡基因组中扩增出含有ＧＨ基因内含子１和４的片段，再用ＭＳＰ１内切酶对产物做ＲＦＬＰｓ分析。由于特异性ＰＣＲ的条件要… 相似文献
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Joan Belcher 《Canadian journal of veterinary research》1954,18(4):120-127
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Influence of Cysteamine on In Vitro Maturation, In Vitro and In Vivo Fertilization of Equine Oocytes
S Deleuze CS Dubois M Caillaud B Bruneau G Goudet G Duchamp 《Reproduction in domestic animals》2010,45(1):1-7
The effect of cysteamine on in vitro nuclear and cytoplasmic maturation of equine oocytes collected by transvaginal ultrasound guided follicular aspiration was assessed. Oocytes were matured in vitro with (cysteamine group) or without (control group) cysteamine. The nuclear stage after DNA Hoechst staining, penetration rates after two different in vitro fertilization (IVF) techniques (IVF media with ionophore and Hepes buffer with heparin) and the embryo yield following oocyte intra-oviductal transfer were used as a criterion for assessing nuclear and cytoplasmic maturation, respectively. Contrary to the data described in other domestic species, there was no effect of cysteamine on in vitro nuclear maturation, IVF or in vivo embryonic development under our conditions. Ovum pick up yields (52%) and maturation rates (control group: 47% and cysteamine group: 55%) were similar to those previously reported. From 57 oocytes transferred to the oviduct in each group, the number of embryos collected was 10 (17%) in the control group and five in the cysteamine group (9%). Those two percentages were not statistically different (p > 0.05). No effect of IVF technique was seen on the success rate (6%) in each group. 相似文献
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S Apichela MA Jiménez‐Díaz M Roldan‐Olarte JN Valz‐Gianinet DC Miceli 《Reproduction in domestic animals》2009,44(6):943-951
Sperm reservoirs in South American Camelids would be crucial for successful fertilization. Since ovulation occurs approximately 36 h after mating, the maintenance of the sperm viability in the oviduct waiting for the ovum is a critical reproductive event. Our study aimed at determining whether the isthmus or the utero tubal junction (UTJ) could function as a sperm reservoir in llama by means of in vivo and in vitro experiments. For the in vivo experiments, the oviducts of adult females with a dominant follicle bigger than 7 mm were examined for the presence of sperm at 6, 18, 24, 28 and 35 h after mating. The results using scanning and transmission electron microscopy showed ultrastructural differences between isthmus and UTJ with respect to (1) predominance of secretory cells in the UTJ and ciliated cells in the isthmus epithelium and (2) cytoplasmic bulbous projection of the secretory cells in the UTJ. Sperm adhered by a mucus‐like substance were seen only in the UTJ at 6, 18, 24 and 28 h post‐mating. Lack of sperm adhered to oviductal mucosa was observed around ovulation (35 h). In vitro experiments demonstrated higher ability of UTJ epithelial cell explants with respect to isthmus explants to bind sperm in a co‐cultured system. The anatomical features and the presence of a sperm bonding agent in the UTJ together with the in vitro differential binding of sperm to UTJ explants strongly suggest that both may be feasible mechanisms that facilitate sperm storage in this oviductal region in llama. 相似文献
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The in vitro tests utilized to evaluate sperm quality represent an interesting and important approach to evaluate ejaculated fecundant capacity. In the present work, the oocyte-penetrating ability of sperm from two bulls with low and two with high in vivo fertility rate was investigated. Sperm-quality parameters, such as sperm concentration, total and progressive motility, acrosome and total sperm anomalies, proximal cytoplasmic drops and live : dead sperm ratio were assessed and batches of sperm homologues in these parameters were selected. In expt 1, a sperm : oocyte ratio of 3000 was used and the oocytes were fixed 5, 10, 15 and 18 h post-insemination (hpi). In expt 2, the sperm : oocyte ratio was reduced to 1000 and the eggs were fixed at 8 and 10 hpi. The results, analysed by chi-square test, showed significant differences (p <0.001) among bulls at 15 hpi in expt 1 and 8 hpi in expt 2; nevertheless, no accordance was found between sperm penetration rate and in vivo fertility. At 18 hpi, the low-fertility bulls exceeded the two high-fertility bulls, supporting previous reports that suggest an opposite correlation between in vivo and in vitro fertility rate at low doses of heparin. Furthermore, a more efficient zona binding ability of one high-fertility bull was pointed out in expt 2 after the reduction of sperm : oocyte ratio, as it reached the highest percentage of penetration when compared with all the others. 相似文献