花鲈胚胎干细胞移植及嵌合体的构建

旨在通过花鲈胚胎干细胞(LJES1)移植构建嵌合体,证实LJES1细胞的体内分化能力和发育全能性.采用脂质体介导法将线性化pCMV-EGFP质粒导入到长期培养的LJES1细胞内,细胞绿色荧光蛋白(GFP)的表达率为5%～10%,以此为供体,通过显微注射方法把20～50个LJES1细胞移植到花鲈囊胚中,共移植囊胚478枚,获得了20枚表达GFP的嵌合体胚胎,其中的15枚胚胎发育成鱼苗.荧光显微观察和PCR检测显示了LJES1细胞可在宿主胚胎内片状嵌合和单细胞分散嵌合,嵌合部位可分布于胚体的三个胚层;同时也证明GFP是一种优良的遗传标记.本实验结果为证实LJES1细胞的发育全能性提供了有力证据.     

Optimal conditions for mycelial growth of Schizosaccharomyces japonicus cells in liquid medium: it enables the molecular investigation of dimorphism

The non‐pathogenic dimorphic fission yeast, Schizosaccharomyces japonicus, could be a suitable model organism for investigation of the genetic background of mycelial growth, as it has a haploid chromosome set and its genome is sequenced. Since earlier results have suggested that its morphological transition required solid substrates, but molecular biological experiments would require hyphae production in a liquid medium, we wanted to find circumstances which would enable hyphae production in liquid media. Several external conditions were investigated, but the strongest inducer was fetal bovine serum (FBS). Its positive effect could be hampered by heat and was dependent on pH, temperature and concentration of the serum. Other protein‐containing compounds, such as peptone and bovine serum albumin or amino acids, proved to be ineffective or weak. Generally, the uninduced and induced mycelial growth of Sz. japonicus could be improved by lower external pH and higher temperature. Copyright © 2014 John Wiley & Sons, Ltd.     

甲嘧磺隆对两种滩涂动物的急性致毒试验

本文生要研究了甲嘧磺隆对鲈鱼、蛏两种主要的滩涂养殖生物进行了急性毒性研究,为甲嘧磺隆用于滩涂防治互花米草提供科学依据。结果表明：处理96h后对于鲈鱼和蛏的半致死浓度（LC50）分别为151．322和374．450mg／L,甲嘧磺隆对两种实验动物急性毒性属于低毒,对于两种滩涂养殖动物在急性毒性方面没有明显影响。     

Novel episomal vectors and a highly efficient transformation procedure for the fission yeast Schizosaccharomyces japonicus

Schizosaccharomyces japonicus is a fission yeast for which new genetic tools have recently been developed. Here, we report novel plasmid vectors with high transformation efficiency and an electroporation method for Sz. japonicus. We isolated 44 replicating segments from 12 166 transformants of Sz. japonicus genomic fragments and found a chromosomal fragment, RS1, as a new replicating sequence that conferred high transformation activity to Sz. japonicus cells. This sequence was cloned into a pUC19 vector with ura4⁺ of Sz. pombe (pSJU11) or the kan gene on the kanMX6 module (pSJK11) as selection markers. These plasmids transformed Sz. japonicus cells in the early‐log phase by electroporation at a frequency of 123 cfu/µg for pSJK11 and 301 cfu/µg for pSJU11, which were higher than previously reported autonomously replicating sequences. Although a portion of plasmids remained in host cells by integration into the chromosome via RS1 segment, the plasmids could be recovered from transformants. The plasmid copy number was estimated to be 1.88 copies per cell by Southern blot analysis using a Sz. pombe ura4⁺ probe. The plasmid containing ade6⁺ suppressed the auxotrophic growth of the ade6‐domE mutant, indicating that the plasmid would be useful for suppressor screening and complementation assays in Sz. japonicus. Furthermore, pSJU11 transformed Sz. pombe cells with the same frequency as the pREP2 plasmid. This study is a report to demonstrate practical use of episomal plasmid vectors for genetic research in Sz. japonicus. RS1 has been submitted to the DDBJ/EMBL/GenBank database (Accession No. AB547343). Copyright © 2010 John Wiley & Sons, Ltd.     

提取麦冬皂苷用酶的微生物筛选及其产酶条件

为提高麦冬总皂苷提取率,采用生物酶法提取麦冬总皂苷,筛选得到产酶微生物Absidiasp.O8s菌株.通过实验确定了该菌产酶的最适条件:诱导物为豆粕,诱导物浸出液在培养基中的体积分数为15%,发酵温度30℃,培养时间为5 d.     

盐水超冷却处理对冰藏鱼品质的影响研究

研究了盐水超冷却预处理对鲈鱼冰藏的品质影响,以-6℃冰水混合物(含盐量6.0%)为冷却介质,对鲈鱼分别预冷却0、40、60、100min,冰藏15d。研究结果表明,超冷却预处理60min和100min的实验组,菌落总数较其它实验组显著下降(p<0.05),Ca2+-ATPase比活力较高,而K值上升较慢。综合考虑储藏过程中生化指标的变化,以及预处理的能耗和时间,超冷却预处理60min对鱼体的储藏保鲜具有较好的指导价值。     

刺参多糖急性毒理研究及其安全性评价

以Wistar大鼠及昆明种小鼠为实验动物，考察了刺参多糖急性毒理作用及安全性。结果表明，刺参多糖属实际无毒物质，可安全地作为保健食品辅助刺或食品添加剂使用。     

超声波提取对麦冬多糖结构和抗氧化活性的影响

研究了超声波提取法在提取麦冬多糖过程中,对麦冬多糖的结构和抗氧化活性的影响.利用气相色谱法、环境扫描电子显微镜、原子力显微镜对其进行初步结构分析,然后在体外化学模拟条件下,测定它的总还原能力、对羟基自由基的清除作用及对Fe2诱发的脂质过氧化反应的抑制作用,并与传统热水提取法提取得到的麦冬多糖进行对比.超声波提取得到的麦冬多糖(SBP)和传统热水提取法提取得到的麦冬多糖(WBP)具有相同的单糖组成,都包括阿拉伯糖、甘露糖、葡萄糖及半乳糖,环境扫描电子显微镜的结果显示经超声提取的多糖的片状结构有不同程度的变小;原子力显微镜的扫描分析显示超声波使其糖链的分子片断变小,且SBP的抗氧化活性都要高于WBP.经超声波处理不会影响多糖的单糖组成,但会改变多糖的微观结构同时提高多糖的抗氧化活性.     

基于高通量测序分析不同保鲜冰处理对鲈鱼菌群组成与代谢功能的影响

目的：分析不同保鲜冰处理下鲈鱼微生物菌群变化及其与品质指标的相关性。方法：分别采用流化冰（slurry ice,SI）、酸性电解水冰（acidic electrolyzed water ice,AEWI）和碎冰（crushed ice,CI）对鲈鱼进行处理,测定鲈鱼的总挥发性盐基氮（total volatile basic nitrogen,TVB-N）含量与K值,并利用高通量测序技术对样品菌群结构和代谢功能进行比较分析。结果：SI和AEWI对鲈鱼均有一定的保鲜作用,其中SI抑制鲈鱼蛋白质和核苷酸降解速度的效果最显著。在冰藏前期（0～3 d）,AEWI与CI组样品中芽孢杆菌属（Bacillus）、乳球菌属（Lactococcus）、大洋芽孢杆菌属（Oceanobacillus）相对丰度明显高于SI组。冰藏中、后期（12～21 d）,SI组样品中假单胞菌属（Pseudomonas）、希瓦氏菌属（Shewanella）、摩替亚氏菌属（Moritella）、埃希氏杆菌属（Escherichia）和别弧菌属（Aliivibrio）相对丰度明显增加,AEWI组样品假单胞菌属（Pseudomonas）和不动杆菌属（Acinetobacter）相对丰度所占比例逐渐增大,CI组中不动杆菌属在21 d时达27.77%,成为造成鲈鱼贮藏中、后期腐败的主要菌属。主成分分析表明,冰藏方式及贮藏时间对鲈鱼菌群结构组成有较大影响。鲈鱼菌群中与TVB-N含量、K值呈正相关的有假单胞菌属、不动杆菌属、嗜冷杆菌属（Psychrobacter）和希瓦氏菌属,它们可能是鲈鱼冰藏期间的优势菌。CI和AEWI组鲈鱼细菌中参与氨基酸、脂质和碳水化合物代谢的相关基因相对丰度高于同一时期的SI组,这在一定程度上从细菌代谢水平解释了SI能延缓鲈鱼腐败的原因,为后期进一步从微生物抑制机制上优化鲈鱼贮藏保鲜技术提供了理论参考。