Vitellogenin in tilapia (Oreochromis mossambicus): Induction of two forms by estradiol,quantification in plasma and characterization in oocyte extract

Two forms of vitellogenin were isolated by DEAE agarose ion-exchange chromatography from plasma of the tilapia, Oreochromis mossambicus. The monomers have apparent molecular masses of 200 and 130 kDa, as indicated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and a total amount of phosphorus of 1.7 and 0.1%, respectively. Antibodies specific to the two forms, designated tVTG-200 and tVTG-130, were generated in rabbits and used to develop enzyme-linked immunosorbent assays (ELISAs) and in Western blot analyses of plasma and oocyte extract. SDS-PAGE of the oocyte extract showed a major protein band at 106.6, minor bands at 26.6, 24.2, and 23.7 kDa, and very faint bands at 83.4 and 17.5 kDa. Western blots of the oocyte extract revealed that the antiserum to tVTG-200 recognized strongly the protein bands at 24.2 and 23.7 kDa, and less strongly the bands at 25.1 and 22.6 kDa, whereas the antiserum to tVTG-130 recognized mainly the protein band at 106.6 kDa. The presence of both VTGs in untreated male tilapia was detected with the ELISAs using relatively high plasma volumes. Their presence in males was confirmed by VTG-like immunoreactive materials eluting from the ion-exchange column at the same positions as tVTG-200 and tVTG-130. The concentrations of the VTGs in males were several orders of magnitude lower than in vitellogenic females. Treatment of male tilapia with estradiol-17β (E₂) induced both VTGs within 24h. After 7 days, tVTG-130 reached a maximum concentration in plasma, whereas tVTG-200 continued to increase. Our findings demonstrate that the two vitellogenins are biochemically distinct, possibly differentially regulated, and made by both sexes.     

条斑紫菜HSP90基因的克隆与表达分析

为了研究HSP90在条斑紫菜胁迫耐受中的作用,克隆了条斑紫菜HSP90基因(命名为PyHSP90),采用定量RT-PCR研究了其表达规律。PyHSP90基因包含一个长2274nt的完整开放阅读框,编码757个氨基酸。PyHSP90蛋白定位于细胞质中,具有HSP90蛋白家族的特征序列和保守结构域,与多种生物的HSP90具有较高的序列一致性。在进化树上条斑紫菜等红藻的HSP90与隐藻的亲缘关系最近,与绿藻和陆生植物亲缘关系较远。低温和高温胁迫都能显著性地诱导条斑紫菜叶状体PyHSP90基因的表达,而且表达量与胁迫程度成正相关;低盐胁迫下PyHSP90基因表达上调;而失水胁迫下PyHSP90基因表达下调。     

饲料中动物源性成分显微近红外光谱检测分析

探讨了应用显微近红外光谱分析技术检测饲料中动物源性成分的可行性.利用12个动物源性成分饲料样品和14个植物源性成分饲料样品建立偏最小二乘定标模型,数学预处理方法为导数+多元散射校正,利用平均光谱来优化模型实现数据压缩,模型的决定系数、交互验证标准差分别为0.969、0.090.4个配合饲料样品作为外部验证未出现误判.结果表明显微近红外光谱分析技术可用于检测饲料中动物源性成分.     

病程相关蛋白与植物抗病性关系的研究及其在草坪草抗病育种中的应用

植物被各类病原物侵染后,发生一系列的生理生化变化,包括细胞壁变厚,产生植保素以及防御相关蛋白。本研究从病程相关蛋白编码基因的诱导和表达、在植物组织细胞中的分布和运输、生物学功能与抗病性的关系、转基因植物的研究等方面依次进行总结,并对草坪病害的研究进展以及病程相关蛋白在草坪草等植物抗病育种工作中的作用与前景进行了讨论。关键词：病程相关蛋白;植物抗病性;病害;抗病育种;草坪草     

热处理对肉类蛋白质构的影响

加热作为最传统的肉熟化和杀菌手段,在今后的水产及畜禽肉加工产业中将依旧占主导地位。作者简述了肌肉蛋白的组成和结构,讨论了不同来源肌肉蛋白在加热过程中的变性行为。水生及陆生来源的肌肉中,大部分肌浆蛋白在40℃~60℃之间聚集,但对一些肌浆蛋白来说,加热到90℃才凝固。溶液中肌原纤维蛋白在30℃~32℃开始展开,在36℃~40℃蛋白质—蛋白质开始结合,在45℃~50℃产生凝胶化(conc.>0.5重量%)。在53℃~63℃的温度下,胶原蛋白开始变性,随后胶原纤维发生收缩。如果胶原纤维不能通过耐热分子间键稳定,则它在进一步加热时溶解并形成胶状。此外,作者还概述了不同种类肌肉蛋白在加热过程中的结构变化及其对质地品质的影响。对于一般完整肌肉的肉块,一定温度的热处理后,显微组织收缩,肌节长度缩短,持水性降低,韧性发生1次或者2次的跃升。鱼糜制品、汉堡肉饼及乳化香肠等碎肉产品,因加工工艺影响,肌肉系统的结构大幅改变。鱼糜制品形成过程较畜禽肌肉复杂;鱼类肌肉中盐溶性蛋白在加热作用下通过分子间二硫键、疏水作用力、氢键等等的作用,包裹水分重新形成立体网状结构,成为具有凝胶特性的鱼糜制品。通常畜禽类肌肉没有鱼糜凝胶形成的能力强。与乳化香肠相比,汉堡肉整体纤维和纤维碎片的出现率更高,其含量高达50%~70%,导致受热后收缩程度更大。乳化香肠中,因肌肉彻底粉碎搅拌因而肌原纤维蛋白被抽提出,在加热时产生密集的蛋白质网络—凝胶,通过毛细管作用力可有效地保持水分。了解不同种类肉蛋白质在烹饪过程中的结构变化以及质构品质变化机理,可为制定更合理的肉类加工方式提供理论依据。     

双斑东方鲀三倍体育苗生产性研究

采用温度休克方法诱导双斑东方鲀三倍体,在水温18-20℃条件下,卵受精后5分钟用40℃的水处理双斑东方鲀受精卵10分钟,三倍体诱导率100％,孵化率相对诱导量达25％;育苗采用前期室内工厂化培育与后期土池培育相结合方法,培育出全为三倍体(体长3 cm以上)苗种5．8万尾,对应孵出苗量,成活率为34．6％。     

Culture of mantle epithelial cells expressing shell matrix proteins from scallop Patinopecten yessoensis

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.     

Intra- and interspecific phenotypic characteristics of fish-pathogenic Edwardsiella ictaluri and E. tarda

Intra‐ and interspecific characteristics of fish‐pathogenic Edwardsiella ictaluri, and E. tarda were determined by numerical analysis of gel electrophoresed protein profiles, fatty acid methyl esters (FAMEs) and immunoblotting. The 18 E. ictaluri isolates revealed a high degree of homogeneity (70% similarity or higher) in their protein profiles and 95% similarity in their FAME, while the nine E. tarda isolates revealed 30% similarity in their protein profiles and 95% similarity in their FAME. Immunoblots probed for antigenic epitopes with goat antiserum produced against E. ictaluri and E. tarda, respectively, revealed that E. ictaluri were more homogeneous compared with the E. tarda isolates. Overall, there was a considerable degree of relatedness between the two species. Our findings suggest that phenotypically E. ictaluri represents a clonal bacterial population structure compared with the less monomorphic E. tarda.     

Temperature modeling of a land-based aquaculture system for the production of Gracilaria pacifica: Possible system modifications to conserve heat and extend the growing season

Temperature control is a major cost for numerous aquaculture systems. Solar thermal engineering techniques can be used to identify inexpensive methods for conserving and capturing heat. Gracilaria pacifica, also known as the culinary ingredient ogo, is currently grown in land-based tanks at a site in Goleta, CA where influent sea water temperatures infrequently reach the 21–28 °C range that provides for optimal growth. The major objective of this study was to explore various designs of a G. pacifica tank culture system that maintain optimal water temperature year round to maximize growth. A model was constructed and calibrated by comparing results to a one-third scale pilot system operated in Davis, CA. For model calibration the most sensitive parameter such as cover optical properties were adjusted first and less sensitive parameters were adjusted later. The pilot system consisted of six tanks, three insulated with foam and a clear polyethylene cover (experimental), and three uninsulated and uncovered (controls). The model had weather data inputs including air temperature, humidity, wind speed, and solar radiation. The model was then compared to a full-scale system operated in Santa Barbara during the winter. The experimental pilot system was 4.93 °C warmer than the control pilot system under optimal weather conditions. The full-scale experimental system was 2.80 °C warmer than the control system under non-ideal conditions. The model demonstrated predictive accuracy under most weather conditions. Furthermore the model is robust enough to accept estimated values for many inputs and still produce accurate results, this suggests a simpler model may be feasible. A polyethylene cover and insulation are not sufficient in general for raising the water temperature to the optimum range during the winter; they may be during other times of the year when more solar energy is available, thereby extending the growing season.     

脊尾白虾热休克蛋白HSP90基因的原核表达与鉴定

将脊尾白虾热休克蛋白90基因克隆到原核表达载体pET-30a中,经酶切验证和DNA测序鉴定后,将重组质粒转化表达宿主大肠杆菌Rosetta,优化温度、时间、IPTG和OD600表达条件进行诱导表达,收集菌液,进行SDS-PAGE和质谱检测,并用Quantity one软件分析蛋白表达水平。结果表明,成功构建了含脊尾白虾HSP90基因的重组表达载体pET-30a-HSP90,表达目的蛋白相对分子量为82.7kD,为HSP90蛋白。通过条件优化认为重组菌株Rosetta/pET-30a-HSP90的最佳诱导温度为37℃,最佳IPTG浓度为1.0mmol/L,最佳诱导时机和诱导时间分别为0.58h、7h。