大鲵α1-血红蛋白基因的生物信息学分析及组织表达研究

 从大鲵皮肤cDNA文库中分离获得大鲵α1 血红蛋白基因全长cDNA序列,生物信息学分析表明,大鲵α 血红蛋白基因全长cDNA序列为594 bp,开放阅读框共432 bp,编码144个氨基酸,编码的大鲵α1 血红蛋白推测的理论分子量为16048.3 u,等电点为6.44。氨基酸组成中酸性氨基酸11.9%,中性氨基酸69.9%,碱性氨基酸18.2%。结构分析表明,该蛋白没有信号肽,但在100~122个氨基酸处以及98~122个氨基酸处分别有由里向外、由外向里的跨膜螺旋区,二级结构以α 螺旋为主。氨基酸序进行同源性列比对表明与人、猕猴的亲缘关系最近,和牛蛙的亲缘关系最低只有35%。荧光定量PCR结果表明,α1 血红蛋白基因在肌肉中表达量最高,在胃中最低。     

黄颡鱼雌、雄个体血清蛋白、酯酶同工酶及血红蛋白的电泳分析

采用聚丙烯酰胺梯度凝胶垂直板(PAGE)电泳,对黄颡鱼雌、雄个体血清蛋白、酯酶同工酶(EST同工酶)及血红蛋白(Hb)进行了分析。结果表明：黄颡鱼雌、雄个体的血清蛋白存在明显差异,表现在B1区雄性个体比雌性个体多1条MI带,B2区雌性个体比雄性个体多1条MA带,少2条MⅡ带,B3区雄性个体比雌性个体多1条MA带。另外在B2区雌、雄个体间MA带和MI带排列次序也有差异。血清EST雌、雄个体也有明显的差异,雄性黄颡鱼血清EST的酶谱出现了3条酶带无中强带,而雌性黄颡鱼EST的酶谱出现了7条酶带,有4条中强带。雌、雄个体胁只能分离出1条谱带,并且雌、雄之间基本没有差异。     

暗纹东方鲀冻藏品质劣化的原因解析

通过比较冰晶、内源蛋白酶以及氧化对冻藏暗纹东方鲀(Takifugu obscurus)品质的影响,确定冻藏暗纹东方鲀品质劣化的关键因素。实验通过液氮浸渍快速冻结抑制鱼肉组织中冰晶增长、碘乙酸浸泡前处理抑制鱼肉内源蛋白酶活性、复合抗氧化剂浸泡前处理抑制蛋白/脂质氧化作用的设计,区分冰晶、内源蛋白酶和氧化3种因素对冻藏鱼肉品质的影响;并以硬度、解冻流失率、蒸煮损失率、挥发性盐基氮含量、K值、Ca2+-ATPase活性、肌原纤维小片化指数和肌纤维长度作为鱼肉品质的评价指标。结果显示,冻结过程中控制冰晶增长对暗纹东方鲀冻藏品质的改善作用最大,冻藏24周后,速冻组的硬度比内源蛋白酶抑制组和氧化抑制组分别提高了26.8%和20.5%,解冻流失率分别降低了44.2%和44.8%,且该处理组TVB-N、K值和Ca2+-ATPase活性变化更慢,24周后分别为10.5 mg/100 g、6.8%和1.43 μmol Pi/mg/10 min。综合各项指标发现,冰晶、内源蛋白酶和氧化对冻藏暗纹东方鲀品质劣化均有影响;其中,冰晶作用是导致暗纹东方鲀在冻藏过程中品质下降的最主要影响因素,内源蛋白酶和氧化对冻藏暗纹东方鲀品质的影响次之,且二者对品质影响无显著性差异。     

Mechanism and Progress About Protein Crystal Growth by Atomic Force Microscopy

Atomic Force Microscopy(AFM) plays a very important role in researching crystal growth mechanism, especially protein crystals. The principle of AFM for protein crystal examination is briefly described as well as the importance of research on protein crystal growth, known growth mechanisms including 2-D steps nucleation, spiral dislocation, normal growth, 3-D nucleation and anisotropic development, and latest progress about eucommia antifungal protein(EAFP) and Trichosanthin(TCS) protein crystal growth mechanism, respectively. Finally, protein growth under microgravity environment is also expounded here and their development tendency and practical prospect.     

中药复方Ⅱ对鲤鱼耐缺氧能力、RBC及HB的影响

研究了中药复方Ⅱ对鲤鱼耐缺氧能力和红细胞(RBC)数及血红蛋白(HB)含量的影响。选用体格健康的鲤鱼80尾,平均分成4组,即基础饲料组(对照组)和3个基础饲料加中药组(实验组),后3个实验组设了3个中药添加量。用上述4种饲料对实验鱼饲养120 d后,测定鲤鱼密闭性缺氧浮头时间、红细胞数和血红蛋白含量。结果显示:3个剂量的中药复方Ⅱ实验组能不同程度地延长缺氧条件下鲤鱼浮头时间,增加鲤鱼血液中红细胞数和血红蛋白含量,但低剂量组与对照组相比,差异不显著(P>0.05)。本研究表明中药复方Ⅱ能提高鲤鱼耐缺氧能力,并有改善外周血象的作用。     

Hemoglobin and subunit multiplicity in the rainbow trout (Oncorhynchus mykiss) hemoglobin system

Hemoglobin (Hb) multiplicity is a commonly used index of phylogenetic differentiation and molecular adaptation in fish. Despite improvements in technologies for characterizing protein structure, our knowledge of the component composition of the Hb system in rainbow trout, Oncorhychus mykiss, has remained almost unchanged through three decades. The Hb is considered to consist of four components, of which two (HbI and HbIV) have been extensively characterised with regard to structure and function. Using a variety of molecular techniques we demonstrate the presence of at least nine different Hb fractions composed of nine (five α and four β globin) chains, and that HbI and HbIV consist of two and five individual components, respectively. These findings indicate that the published data on trout HbI and HbIV refer to mixtures of isoHbs and need to be reappraised. This revised version was published online in August 2006 with corrections to the Cover Date.     

透明颤菌血红蛋白基因在多粘芽孢杆菌中的克隆和表达

本研究将启动子P43与透明颤菌血红蛋白(Vitreoscilla hemoglobin)基因(vgb)通过重叠PCR进行融合,克隆到芽孢杆菌表达载体p CM20上,将筛选得到的重组载体p CM20-vgb转化多粘芽孢杆菌(Paenibacillus polymyxa)NR1,Western-Blot表明重组菌株表达了VHb蛋白,该蛋白的表达对重组菌体的生长和多粘菌素的产生均有促进作用。     

Mastitis Whey — a Good Medium for Bacteria?

Growth of mastitis pathogenic bacteria was measured in bovine whey samples by a turbidometric microtechnique. Whey from mastitis cows supported growth as compared with whey prepared from normal milk. Blood proteins leak into milk during mastitis. A study was undertaken to analyze which molecules from blood would promote bacterial growth in whey Fractions containing hemoglobin showed a distinct growth-promoting effect. An inadequate iron supply is one of the restricting growth factors for bacteria in milk. By utilizing heme-compounds the pathogens can by-pass the effect of antimicrobial iron-binding present in milk in the form of lactoferrin.