利用盐溶蛋白PAGE电泳法检测玉米种子的研究

应用玉米盐溶蛋白PAGE电泳方法检测了10个玉米品种的纯度及其杂交种的真实性。通过品种间的盐溶蛋白谱带的对比,发现其中有2条蛋白带是各品种所共有的,而除这些共有的蛋白带以外,各品种还具有一些特异条带。根据每个品种特有的蛋白条带,建立其特征条带图谱,可应用于该品种的相关鉴定工作。     

多枝赖草基因组Mb级DNA制备和酶切方法的优化

Mb级DNA的制备和酶切是构建YAC,PAC,BAC,BIBAC和TAC文库与大尺度物理图谱的基础。以浓度0.4%(m/m)NaCl人工模拟盐池中生长的多枝赖草再生叶为材料提取细胞核,经钢丝细胞筛过滤,多次回收滤渣和离心上清中的细胞核,相差显微镜调浓度达108个/mL后,经LMP包埋,蛋白酶K降解核蛋白,脉冲电泳纯化回收后,在LMP胶块中2 Mb以上DNA的浓度约为250 ng/μL。在胶中经Hind III部分酶切再用脉冲电泳回收不同大小范围的DNA,电洗脱浓缩除盐后浓度约为25 ng/μL。用此方法得到的核DNA不但无细胞器DNA等的污染,而且浓度高,可直接用于各种人工染色体文库的构建。     

Genetic and phenotypic comparison of Nocardia seriolae isolated from fish in Japan

The phenotypic and genetic characterizations of 58 isolates of the fish pathogen Nocardia seriolae , from amberjack, Seriolae dumerili , yellowtail, Seriola quinqueradiata , Japanese flounder, Paralichthys olivaceus , and chub mackerel, Scomber japonicus, in Japan from 1970–2005, were examined to investigate the epidemiological relationship between isolates. The phenotypic and genetic characterizations were determined by α-glucosidase activity and biased sinusoidal field gel electrophoresis (BSFGE) analysis, respectively. There was no α-glucosidase activity in strains isolated from 2000–05 ( n  = 50) with a few exceptions ( n  = 3), while all strains isolated from 1970–90 ( n  = 8) were positive. In BSFGE analysis, digestions with restriction enzymes Xba  I and Ase  I produced 15 and 16 restriction patterns, respectively. All restriction patterns obtained from 50 strains isolated during 2000–05 were unrelated to those obtained from eight strains isolated during 1970–90, with the exception of two strains isolated during recent outbreaks. Based on the phenotypic and genetic characterizations, recent outbreaks of nocardiosis in Japan are suggested to be epidemiologically unrelated to earlier outbreaks in Japan. Although a low genetic relationship was observed in the restriction pattern between recent and earlier isolates, identity was confirmed between these groups of isolates because five representative strains showed 99.9% homology with N. seriolae ATCC43993^T in the 16S rRNA sequence.     

兴国红鲤、草鱼及其杂种一代血清蛋白质的电泳分析

用聚丙烯酰胺凝胶垂直平板电泳分析了兴国红鲤、草鱼及其杂种一代的血清蛋白。结果表明,它们各有不同的电泳图谱,杂种一代为双亲的嵌合型,相对而言,MA带偏向母本,MI带偏向父本。     

Characterization of Lancefield group C Streptococcus dysgalactiae isolated from farmed fish

A Lancefield group C streptococcal (GCS) infection caused by Streptococcus dysgalactiae that is characterized by severe necrotic lesions of the caudal peduncle has been an increasing cause of mortality in farmed fish such as amberjack, Seriola dumerili, and yellowtail, Seriola quinqueradiata, in the southern part of Kyushu, Japan. In this study, enzymatic profiles of GCS strains from fish and mammals were investigated using the API ZYM system, and genotypic characterization of GCS strains was performed using biased sinusoidal field gel electrophoresis (BSFGE). The partial sequence of the 16S-23S rDNA intergenic spacer region of the GCS strain isolates from fish and mammals was also compared. The API ZYM test indicated that it is difficult to differentiate isolates of S. dysgalactiae from fish and animals based on enzymological variations. In the BSFGE analysis, the macrorestriction profiles, which were obtained using SmaI or ApaI as a restriction enzyme, revealed variations between the fish and animal isolates. The partial sequence of the 16S-23S rDNA intergenic spacer region of all the tested fish isolates differed from all mammalian isolates in one or two nucleotides. The possibility of a clonal expansion of S. dysgalactiae strains in farmed fish was also suggested by the BSFGE profiles of fish isolates.     

Cryoprotective effects of shrimp head protein hydrolysate on gel forming ability and protein denaturation of lizardfish surimi during frozen storage

ABSTRACT:   The effects of shrimp head protein hydrolysate (SHPH) from three species of shrimp (northern pink shrimp [ Pandalus eous ], endeavour shrimp [ Metapenaeus endeavouri ], black tiger shrimp [ Penaeus monodon ]) on gel forming ability and protein denaturation of lizardfish surimi during frozen storage at −25°C were evaluated. The quality of lizardfish surimi with 5% (dried matter) of any of the three SHPH or sodium glutamate (Na-Glu) was examined in terms of gel strength, whiteness, Ca-ATPase activity and the amount of unfrozen water, comparing with those of surimi without additive as the control. The residual Ca-ATPase activity and gel strength of surimi with SHPH were higher than those of the control throughout 180 days of frozen storage, regardless of shrimp species. The highest effect was found in surimi with Na-Glu. The gel strength and Ca-ATPase activity found a high positive correlation. The addition of SHPH to surimi also increased the amount of unfrozen water by approximately 1.29–1.36 fold higher than the control, however kamaboko gels of the control was significantly whiter. From these results, freeze-induced denaturation of lizardfish muscle protein could be lessened by the addition of SHPH, resulting in a high gel strength and Ca-ATPase activity.     

不同沉淀剂对酚抽法提取植物蛋白质效果比较分析

酚抽法已经逐渐成为植物蛋白质组研究中通用的蛋白质提取方法,但在各种改进酚抽法中,应用到的沉淀试剂各不相同,致使蛋白沉淀得率、纯度和沉淀时间也各异,本研究以热带植物橡胶树叶片、胶乳和海马齿叶片为研究材料,在过饱和硫酸铵甲醇溶液、醋酸氨甲醇溶液和丙酮溶液沉淀剂下对比蛋白提取的效果,通过单向电泳检测、胶图质量分析、质谱鉴定、蛋白沉淀时间和得率比较,发现3种沉淀剂提取的蛋白样品1-DE图谱显示的蛋白条带数量均较多,但醋酸铵沉淀法提取蛋白图谱有条纹不清晰现象。得出结论：丙酮沉淀法对橡胶树叶片和胶乳蛋白质的提取率较高,硫酸铵沉淀法对海马齿叶片蛋白质提取率较高。过饱和硫酸铵甲醇溶液和丙酮沉淀剂提取的蛋白质质量较高,所得蛋白图谱背景清晰,质谱鉴定蛋白质信息量大。研究结果可优化提取高质量植物蛋白质的方法,并有望为顽拗类植物蛋白质组学研究提供参考。     

葵花粕中分离蛋白的成分及特性

该文以水酶法提取葵花籽油后的葵花粕为原料,通过盐提酸沉法提取其中的分离蛋白,并对其进行组分分析、氨基酸成分分析、电泳图谱分析和凝胶色谱分析,同时对其溶解性、吸水性、吸油性、乳化性和起泡性等功能特性进行研究。结果显示葵花籽分离蛋白的氨基酸组成与FAO必须氨基酸需要量模式相比赖氨酸含量较低,功能特性好于或接近大豆分离蛋白。葵花分离蛋白含有3个主要组分,其分子量分别为380×10³,100×10³和27×10³,绿原酸与分离蛋白紧密结合。     

新型滴鼻灭活鹦鹉热衣原体疫苗的免疫效力评价

【目的】探究由霍乱弧菌菌影(Vibrio cholerae ghosts, VCG)和纳米壳聚糖凝胶(Gel)组合制作的一种新型灭活衣原体疫苗复合佐剂是否可增强鹦鹉热衣原体原体(elementary body, EB)灭活抗原诱导动物机体产生的免疫应答。【方法】将60只7日龄SPF鸡随机分为4个组,分别为：EB+VCG+Gel组、EB+VCG组、Gel组和EB组。通过滴鼻途径免疫鸡群,免疫2次,每次间隔14 d,免疫后检测鸡血清中IgG抗体水平、淋巴细胞增殖指数、CD4⁺/CD8⁺T细胞比例、细胞因子(白介素-4(IL-4)、IL-10、IL-12和干扰素-γ(IFN-γ))含量及攻毒后鸡喉头排菌量和肺脏的病理损伤程度。【结果】与Gel和EB组相比,EB+VCG+Gel和EB+VCG组IgG抗体水平、淋巴细胞增殖指数、IFN-γ含量和CD4⁺/CD8⁺T细胞比值显著或极显著升高(P<0.05;P<0.01)。此外,与Gel和EB组相比,攻毒后第12天,EB+VCG+Gel组喉头排菌量极显...     

蛋白质组学研究技术与展望

蛋白质组学是继基因组学之后兴起的一门前沿学科。本文简要介绍蛋白质组学的概念和研究意义,以及蛋白质组学的主要研究技术,并展望蛋白质组学的技术前景。