Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction

Vibrio anguillarum , an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum . PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species-specific fragment (689 bp) from V. anguillarum . Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA , or from six colony-forming units (CFU) mL⁻¹ of cultured V. anguillarum . However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g⁻¹ tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species-specific method to detect V. anguillarum in practical situations.     

cDNA cloning and complete primary structures of myosin heavy chains from brushtooth lizardfish Saurida undosquamis and wanieso lizardfish S. wanieso fast skeletal muscles

ABSTRACT:   The complete cDNA sequences encoding predominant types of myosin heavy chain (MYH) in the fast skeletal muscle were determined for brushtooth lizardfish Saurida undosquamis and wanieso lizardfish S. wanieso , which are used as materials for preparing high-quality surimi-based products. The cDNA consisted of 5973 and 5987 bp, respectively, and both encompassed an open reading frame encoding a polypeptide of 1936 amino acid residues. Brushtooth and wanieso lizardfish MYH showed the amino acid sequence identity of 92–93% to white croaker MYH, which was higher than that of 90% to walleye pollack MYH. The putative binding sites for ATP, actin, and regulatory and essential light chains in the subfragment-1 region of brushtooth lizardfish MYH exhibited a high identity with white croaker counterparts as well as the sequences of subfragment-2 and light meromyosin. In contrast, phylogenetic tree, constructed by the neighbor-joining method based on mitochondrial 16S rRNA gene, revealed that the two lizardfish species formed a cluster with walleye pollack, which was paraphyletic with white croaker. Therefore, a good reputation for lizardfish and white croaker to have a high thermal-gel forming ability seemed to be reflected by MYH rather than biological similarity as revealed by the mitochondrial 16S rRNA gene.     

基于食品安全的供应链合作伙伴关系研究

针对国内近年来频繁的食品安全问题,从食品供应链角度进行分析,查找现有食品供应链影响食品安全的隐患,指出构建食品供应链合作伙伴关系对于保障食品安全的重要意义,并提出构建食品供应链合作伙伴关系的措施,更有效保证食品安全。     

农杆菌介导的转高赖氨酸蛋白基因（sb401）水稻T4代分析

通过农杆菌介导的方法用富赖氨酸蛋白基因（sb401）转化粳稻品种日本晴,获得了独立的10个转化株系,对转化株系进行连续自交,通过筛选得到9个纯合的T4代转化株系。 Southern blotting分析发现,整合位点是随机的,并为低拷贝（1~3个）。TAI-PCR扩增得到8个T-DNA侧翼序列,并定位于日本晴的7条染色体上。蛋白质和氨基酸测定分析发现,sb401基因对各株系的蛋白质、赖氨酸和其他氨基酸组分的提高起到了一定的作用。将杂交结果与T-DNA插入位置结合分析发现,在低拷贝的情况下,表达量的差异不明显。     

高温胁迫下水稻胚乳淀粉分支酶各同工型基因的表达特征

 以稻米品质温度敏感型的早籼稻品种浙辐49为材料,利用人工气候箱控温处理试验和实时荧光定量PCR技术,探讨了水稻发育胚乳中淀粉分支酶(starch branching enzyme, SBE)的3种主要同工型基因 (SBEⅠ、 SBEⅢ和SBEⅣ) 在不同温度处理下的相对表达量差异及动态变化特征。结果表明,高温处理对水稻胚乳中SBE基因的表达调控因其同工型的种类而异,SBEⅣ基因在高温处理下呈上调表达,而SBEⅠ和SBEⅢ基因在高温处理下的相对表达量则明显降低,表现为下调表达;与SBEⅣ等同工型相比,水稻胚乳中SBEⅠ基因对高温胁迫的表达响应相对更敏感。     

中国农业面源污染排放的空间差异及其动态演变

以2003—2014年为研究时段,在测算农业面源污染排放强度的基础上,综合运用基尼系数和非参数估计方法,研究我国农业面源污染的空间差异及其动态演变,结果表明:1)我国农业面源污染排放总体下降,且表现出明显的空间差异,东部和中部地区的排放强度较高,西部和东北地区则相对较低。2)2003—2014年,中国农业面源污染排放强度的区域差异略微扩大,地区间差异是其总体差异的主要来源。3)核密度估计结果表明,中国农业面源污染排放总体差异表现为"下降-上升-下降"的波动变化趋势。4)马尔科夫链分析表明,中国农业面源污染在不同类型间的相互流动较为微弱,但从长期来看,存在向两极分化发展的趋势特征。     

油锯人机界面的优化研究

该文通过实验研究探讨了油锯人机界面几何参数对操作者主观施力及对其身体生理负荷的影响规律 .通过对实验数据的回归得出了高把油锯和矮把油锯的人机界面几何参数与评价指标 (最大有效扭矩、力损耗因子、肌肉收缩强度 )之间存在的定量化方程 .建立了油锯的人机界面几何参数优化的目标函数 ,并求出了高把油锯和矮把油锯的最佳几何参数 .高把油锯的最佳几何参数 :把手横轴距为 5 40mm ;把手与地面垂轴距为 6 88mm ;把手与油箱后部纵轴距为 186mm ;把手直径为 30mm ;把手倾角为 33° .矮把油锯的最佳几何参数 :把手横轴距为 14mm ;把手纵轴距为 40 0mm ;把手垂轴距为 5 0mm ;把手直径为 2 8mm ;把手倾角为 2 3     

以“猪疫情”与“猪周期”叠加致肉价波动为契机建立新时代立体生态养殖产业链

该文从2019年中国生猪和猪肉产品价格大幅波动而带动CPI快速上调的“高通胀风险”及与工业生产PPI下降的“高通缩风险”关联性,探讨了推动猪肉价格快速上涨的机制.难控性“猪疫情”、规律性“猪周期”、投机性“猪炒作”及泛化性“外贸战”是促进肉产品价格快速上涨的“启动”、“叠加”、“内讧”及“外推”四大机制,并建议以此为机...     

星斑川鲽MHCⅡ恒定链Ii基因的克隆和表达特性

为了研究星斑川鲽MHCⅡ类分子的作用及调控机制,实验通过SMART-RACE技术克隆得到了星斑川鲽MHCⅡ恒定链(MHCⅡIi)的全长cDNA序列,其长度为1766 bp,包含135 bp的5′非编码区、837 bp的开放阅读框和794 bp的3′非编码区。该基因共编码279个氨基酸。理论分子量为30.848 ku,等电点为6.89。与已知物种MHCⅡIi进行同源性比对,结果与狼鲈、紫红笛鲷和鳜关系较近,同源性均为79%。利用quantitative realtime PCR(qRT-PCR)技术检测了MHCⅡIi在星斑川鲽不同组织中的表达,以及爱德华氏菌感染前后对该基因在不同组织中表达水平的影响,结果显示:在脾脏、头肾、肝脏、后肠、性腺、心脏、血液、鳃和肌肉组织中,MHCⅡIi mRNA均有表达,但在表达量上有明显差异,脾脏和头肾组织相对表达水平较高,鳃、血液、肌肉、心脏和性腺中的表达水平较低。病原感染后,免疫相关组织脾脏和头肾的表达水平升高最明显,肝脏和后肠的表达水平也略有升高,但变化不明显。本研究可为星斑川鲽MHCⅡ类分子的作用机理提供理论依据,同时为海水养殖鱼类的抗病遗传育种工作提供研究基础。