不同生物素水平对3T3-L1脂肪细胞脂肪合成相关基因转录表达的影响

【目的】诱导3T3-L1细胞分化为3T3-L1脂肪细胞,研究不同生物素水平对3T3-L1脂肪细胞脂肪合成相关基因转录表达的影响。【方法】利用三联诱导法将3T3-L1细胞经诱导分化为3T3-L1脂肪细胞。当3T3-L1脂肪细胞密集后分别采用0(对照组)、0.2、0.5、1 μmol/L生物素处理,分别在12 h、24 h与48 h时检测细胞上清液中PK mRNA、GLUT-4 mRNA、FAS mRNA及ACC1 mRNA相对表达量。【结果】在试验12 h时:各试验组GLUT-4、PK、ACC1 mRNA相对表达量均极显著(P＜0.01)高于对照组,1 μmol/L组与0.5 μmol/L组FAS mRNA相对表达量极显著(P＜0.01)高于对照组与0.2 μmol/L组;在试验24 h时:1 μmol/L组GLUT-4 mRNA与PK mRNA相对表达量显著(P＜0.05)高于对照组,1 μmol/L组ACC1 mRNA相对表达量极显著(P＜0.01)高于对照组,0.2 μmol/L组与0.5 μmol/L组ACC1 mRNA相对表达量显著(P＜0.05)高于对照组;在试验48 h时:1 μmol/L组GLUT-4 mRNA相对表达量显著(P＜0.05)高于对照组,1 μmol/L 组PK mRNA相对表达量极显著(P＜0.01)高于对照组,0.5 μmol/L组FAS mRNA相对表达量极显著(P＜0.05)高于1 μmol/L组,1 μmol/L组FAS mRNA相对表达量显著(P＜0.05)高于对照组,1 μmol/L组ACC1 mRNA相对表达量极显著(P＜0.01)高于其它组。【结论】生物素的添加可以提升脂肪细胞GLUT-4、PK、FAS与ACC1 mRNA相对表达量,且当以1 μmol/L浓度作用时对GLUT-4、PK与ACC1提升效果最佳,以0.5 μmol/L浓度作用时对FAS提升效果最佳。     

草鱼基因组中微卫星分子标记的制备及筛选

采用新型的生物素-磁珠吸附微卫星富集法与传统的放射性同位素杂交法相结合的方式,研究了草鱼（Ctenopharyngodon idella）基因组中存在的微卫星资源.结果,从1 000个细菌中,挑出90个阳性克隆,把其中45个进行测序分析,在这些序列中共含有68个微卫星核心序列,有55个含有重复次数大于5次的微卫星核心序列.其中,单一型标记49个,占72%,包括CA重复单元共22个,GT重复单元共18个,其他重复单元为9个;在72%的单一型标记中,完美单一型微卫星标记为44个,占65%.另外,混合型标记共19个,13为完美混合型;在所有微卫星标记中,CA重复类型最普遍,依次为GT、GA等.在68个微卫星标记中,有15个依据引物设计原理设计了引物.     

枯草芽孢杆菌生物素操纵元的初步改造

为了克服生物素生物合成中的抑制作用,提高生物素操纵元基因本底水平的表达,分别构建了整合表达载体pGJj01和pGJj02,通过双交叉整合的方式先后将线性化的pGJj01和pGJj02整合到枯草芽孢杆菌AS1094的染色体上,构建了枯草芽孢杆菌GJZ01,用强启动子P43-1替换掉生物素操纵元自身的启动子Pbiotin,并在bioB和bioI基因间加入强终止子和强启动子P43-2。经PCR和Southern检测表明,整合构建正确。试验可为下一步研究生物素操纵元基因表达提供良好的宿主菌素材。     

Influences of increased levels of biotin, zinc or mannan-oligosaccharides in the diet on foot pad dermatitis in growing turkeys housed on dry and wet litter

Foot pad dermatitis (FPD) is very common in turkeys and it is an animal health and welfare issue affecting not only performance and walking ability but also the carcass quality. Thus, there is a great need to find out the preventive measures against this problem. The potential roles of extra dietary biotin, Zn and mannan-oligosaccharides (MOS) in preventing the development of FPD were assessed in this study which was conducted on 2-week-old female turkeys over a period of 4 weeks. The birds were allotted to four groups, with 29 each, and housed on dry wood shavings in floor pens. The turkeys were fed a control (300 μg biotin and 50 mg Zn/kg), high biotin (2000 μg/kg), high Zn (150 mg/kg) or MOS (1%) diet. Half of the turkeys in each group were additionally exposed to wet litter (27% DM) for 8 h daily in adjacent separate boxes. Foot pads of the birds were examined on days 0, 7, 14, 21 and 28 and assessed macroscopically and histopathologically for foot pad lesions. High dietary levels of biotin or Zn significantly reduced the severity of FPD on dry litter (score 1 vs. 2 in control) but not on wet litter. However, MOS did not affect the severity of foot pad lesions either on dry or wet litter. In addition, the severity was overall substantially higher on wet (approximately three times) than on dry litter. The present results suggest that adding high levels of biotin or Zn to the diet could reduce the development and severity of FPD on dry litter but without having any preventive effects on wet litter. Finally, the high litter moisture appears to be the major factor resulting in FPD. Therefore, the litter should be maintained dry to minimize the prevalence and severity of FPD in turkeys.     

乳及乳制品中游离生物素分析方法研究进展

生物素是动物机体维持正常生理机能所必需的维生素之一。本文综述了乳制品中游离生物素分析的微生物法、色谱法及质谱法,比较了各种方法的优缺点及操作中的注意事项,阐述了企业如何选择生物素分析方法及未来生物素分析的发展趋势。     

Physiological Changes in the Concentrations of Biotin in the Serum and Milk and in the Physical Properties of the Claw Horn in Holstein Cows

Physiological changes in the concentrations of biotin in the serum and milk and in the physical properties of the claw horn were examined in Holstein cows. A lower concentration of biotin in the serum and a higher concentration of biotin in milk were found during early and late lactation and during the dry period, and a significant (p<0.05) inverse correlation was found between serum and milk biotin concentrations. A high moisture content and a low level of hardness of the claw horn were found during mid-lactation. Our results indicate that change in the serum biotin concentration probably results from the loss of biotin in the milk of cows during each stage of lactation and also confirm that the moisture content and hardness of the claw horn undergo physiological changes.     

生物素的应用研究概况

本文对生物素在提高胴体质量、改善肉质,提高家畜的繁殖性能、生产性能及饲料利用率,标记核酸探针以及发酵工业中的应用进行了概述。     

口蹄疫病毒核酸试纸条检测方法的建立

本研究通过核酸探针与免疫层析相结合的方法,建立了一种简单、敏感、特异的检测口蹄疫病毒的方法。试验利用RT-PCR方法获得口蹄疫病毒3D核苷酸片段,在引物中设计了灵敏度高、特异性好的核酸探针--生物素和地高辛,使扩增产物结合探针。利用胶体金的放大原理将链霉亲和素与金颗粒形成胶体金混合物,从而与RT-PCR产物中的生物素探针结合。硝酸纤维素膜上端标记生物素化山羊抗兔IgG作为指控条带,下端标记抗地高辛抗体以捕获RT-PCR产物中的地高辛探针。组装金标试纸条,检测RT-PCR产物,结果表明该核酸试纸条可以检测到 0.3×10^-3~3×10^-3 μg/μL,敏感性高于琼脂糖凝胶电泳,两种方法的符合率高,核酸试纸条检测方法是一种敏感性高、成本低且费时短的新型检测方法。该方法为口蹄疫病毒检测提供了一种新的思路。     

不同生物素水平对3T3-L1脂肪细胞脂肪分解及相关基因表达的影响

为研究不同生物素水平对3T3-L1脂肪细胞脂肪分解及相关基因表达的影响,本试验利用三联诱导法将3T3-L1细胞经诱导分化为3T3-L1脂肪细胞。当3T3-L1脂肪细胞密集后分别采用0（对照组）、0.2、0.5、1.0 μmol/L生物素处理,分别在12、24与48 h时检测细胞上清液中甘油释放量、脂肪甘油三酯水解酶（ATGL）、激素敏感性脂肪酶（HSL）、脂滴包被蛋白A （perilipin A）及脂肪分化相关蛋白（ADRP）相对表达量。结果显示,在12 h时,1.0 μmol/L组甘油释放量、ATGL基因mRNA相对表达量均极显著低于对照组（P<0.01）,0.5、1.0 μmol/L组HSL基因mRNA相对表达量显著低于对照组（P<0.05）,1.0 μmol/L组perilipin A、ADRP基因mRNA相对表达量均极显著高于对照组与0.2 μmol/L组（P<0.01）;在24 h时,各试验组甘油释放量、1.0 μmol/L组ATGL基因mRNA相对表达量均极显著低于对照组（P<0.01）,0.5、1.0 μmol/L组perilipin A基因mRNA相对表达量极显著高于对照组与0.2 μmol/L组（P<0.01）;1.0 μmol/L组ADRP基因mRNA相对表达量极显著高于其他组（P<0.01）;在48 h时,1.0 μmol/L组甘油释放量、ATGL基因mRNA相对表达量均极显著或显著低于对照组（P<0.01;P<0.05）,1.0 μmol/L组perilipin A基因mRNA相对表达量显著高于对照组与0.2 μmol/L组（P<0.05）,0.5、1.0 μmol/L组ADRP基因mRNA相对表达量极显著高于对照组（P<0.01）。综上所述,生物素可通过促进perilipin A与ADRP表达,同时抑制ATGL与HSL表达,达到抑制脂肪分解的效果,且浓度为1.0 μmol/L时效果最佳。     

鸡生物素缺乏症研究进展

生物素缺乏导致体内三大物质代谢紊乱,从而影响鸡的生长和生产性能。生物素缺乏症的病鸡免疫机能下降,影响肝脏中的丙酮酸羧化酶活性以及血清碱性磷酸酶活性,影响葡萄糖和血清总磷水平,造成神经内分泌功能紊乱等。分析鸡对生物素吸收利用的因素,总结生物素缺乏症的诊断指标和临床症状,以及在饲料中适宜添加量,可为进一步研究该病提供理论依据。