两种氨酸对公牛精液冷冻效果研究

[目的]试验研究L—半胱氨酸、精氨酸对牛精液冷冻效果。[方法]基础液中分别添加不同浓度的L—半胱氨酸、精氨酸进行试验,再用L₉(3₄⁾正交表优化最佳配比,获得不同的稀释液配方,然后再进行对比试验,优选出使用效果最好的牛精液冷冻稀释液。[结果]牛精液冷冻稀释液添加0.125%、0.25%、0.375%的半胱氨酸;牛精液冷冻稀释液添加0.5%、0.75%精氨酸,牛冷冻精液解冻活力均得到提高。[结论] 结果表明,半胱氨酸、精氨酸可用于牛精液冷冻保存,能改善牛精液冷冻保存效果,对提高牛冷冻精液质量效果明显。     

相似穿孔线虫S型半胱氨酸蛋白酶基因克隆与序列分析

根据不同种类线虫编码半胱氨酸蛋白酶的保守序列及植物寄生性线虫的半胱氨酸蛋白酶氨基酸密码子的偏好设计简并引物,通过RACE技术,首次从相似穿孔线虫(Radopholus similis)中克隆得到一个编码S型半胱氨酸蛋白酶基因的cDNA全长,命名为Rs-CPS(GenBank登录号EU659125)。Rs-CPS基因全长为1112bp,编码314个氨基酸,分子量为34.69ku。分析结果显示:Rs-CPS氨基酸序列具有半胱氨酸蛋白酶家族典型的Cys-His-Asn三联体酶催化活性中心,而且N端有1个17个氨基酸残基组成的信号肽。     

寄生虫半胱氨酸蛋白酶研究进展

半胱氨酸蛋白酶(Cysteine protease,CP)是一类在酶的活性中心含有半胱氨酸残基的蛋白水解酶.CP属于木瓜蛋白酶家族,广泛分布于从病毒至脊椎动物的生物体中.对人体半胱氨酸蛋白酶的研究主要集中在细胞凋亡分子机制和恶性肿瘤的基因诊断与治疗等方面.而对寄生虫半胱氨酸蛋白酶研究的内容则更为广泛,其中包含半胱氨酸蛋...     

Enzyme Bioprospection of Marine-Derived Actinobacteria from the Chilean Coast and New Insight in the Mechanism of Keratin Degradation in Streptomyces sp. G11C

Marine actinobacteria are viewed as a promising source of enzymes with potential technological applications. They contribute to the turnover of complex biopolymers, such as pectin, lignocellulose, chitin, and keratin, being able to secrete a wide variety of extracellular enzymes. Among these, keratinases are a valuable alternative for recycling keratin-rich waste, which is generated in large quantities by the poultry industry. In this work, we explored the biocatalytic potential of 75 marine-derived actinobacterial strains, focusing mainly on the search for keratinases. A major part of the strains secreted industrially important enzymes, such as proteases, lipases, cellulases, amylases, and keratinases. Among these, we identified two streptomycete strains that presented great potential for recycling keratin wastes—Streptomyces sp. CHA1 and Streptomyces sp. G11C. Substrate concentration, incubation temperature, and, to a lesser extent, inoculum size were found to be important parameters that influenced the production of keratinolytic enzymes in both strains. In addition, proteomic analysis of culture broths from Streptomyces sp. G11C on turkey feathers showed a high abundance and diversity of peptidases, belonging mainly to the serine and metallo-superfamilies. Two proteases from families S08 and M06 were highly expressed. These results contributed to elucidate the mechanism of keratin degradation mediated by streptomycetes.     

Identification of a Group of Novel y-Gliadin Genes

γ-Gliadins are an important component of wheat seed storage proteins. Four novel γ-gliadin genes （Gli-ngl to Gli-ng4） were cloned from wheat （Triticum aestivum） and Aegilops species. The novel γ-gliadins were much smaller in molecular size when compared to the typical γ-gliadins, which was caused by deletion of the non-repetitive domain, glutamine-rich region, 3＂ part of the repetitive domain, and 5＇ part of the C-terminal, possibly due to illegitimate recombination between the repetitive domain and the C-terminal. As a result, Gli-ngl and Gli-ng4 only contained two and three cysteine residues, respectively. Gli-ngl, as the representative of novel γ-gliadin genes, has been sub-cloned into an Escherichia coli expression system. SDS- PAGE indicated that the both cysteine residues of Gli-ngl could participate in the formation of intermolecular disulphide bonds in vitro. Successful cloning of Gli-ngl from seed cDNA of T. aestivum cv. Chinese Spring suggested that these novel γ-gliadin genes were normally transcribed during the development of seeds. Phylogenic analysis indicated that the four novel γ-gliadin genes had a closer relationship with those from the B （S） genome of wheat.     

雌二醇和孕酮对小鼠子宫半胱亚磺酸脱羧酶基因表达和牛磺酸含量的调控

实验主要研究了17β-雌二醇(E2)和孕酮(P4)对小鼠子宫组织中半胱亚磺酸脱羧酶基因(CSD)表达和牛磺酸含量的调控。给去卵巢小鼠分别皮下注射植物油(对照组)、17β-雌二醇(E2)、孕酮(P4)、E2和P4、E2和氟维司群(ICI 182780)、P4和米非司酮(RU486),24h后观察各组小鼠子宫中CSD的表达和牛磺酸的含量。Real-time PCR和Western blot定量分析结果显示,每只小鼠注射1μg E2能够显著抑制CSD的mRNA和蛋白的表达,6mg P4能够显著促进CSD mRNA和蛋白的表达,其受体抑制剂RU486和ICI182780都能逆转它们的作用。高效液相色谱(HPLC)测定结果显示,注射1μg E2和6mg P4分别下调和上调子宫组织中牛磺酸的含量。上述结果提示E2和P4可能通过受体途径参与调节子宫CSD的表达,从而调节子宫中牛磺酸含量。     

Proteases of Clostridium botulinum. V. Studies on the serological relationship between proteases from Clostridium botulinum and other spore-forming bacteria

By the use of the electrophoretic casein precipitating inhibition test (CPI-test) the serological relationship between proteolytic enzymes produced by different species within the genera Clostridium and Bacillus has been tested. The proteases produced by Clostridium botulinum types A, B, C, D and F cross-reacted with each other. Clostridium botulinum strain 84 was inhibited by antiproteases produced against Clostridium sporogenes, Clostridium botulinum types C and F (protease F I and F II), but not by antiproteases against Clostridium botulinum types B and F (protease II), Clostridium bifermentans and Clostridium perfringens. The protease of the newly described Clostridium botulinum strain 89 (type G) was inhibited by Clostridium sporogenes antiprotease, but not by any of the other antiproteases. It is not possible to differentiate between Clostridium botulinum, Clostridium sporogenes and Clostridium perfringens by use of serological differentiation of their proteolytic enzymes. The protease of Clostridium bifermentans is not serologically related to any of the species tested in this investigation. Proteases produced by different Bacilli were not inhibited by antiproteases from Clostridium botulinum types B, C and F, Clostridium sporogenes, Clostridium bifermentans, and the two strains of Clostridium perfringens tested. This investigation indicates a serological relationship between proteases from different Clostridium species, but not a serological relationship between proteases produced by the Clostridium species and Bacillus species tested.     

腐蹄病节瘤拟杆菌的分子致病机制的研究进展

腐蹄病是反刍动物绵羊、山羊、鹿和奶牛等常见的一种高度接触性传染病。节瘤拟杆菌是致病作用的主要病菌之一,它是通过IV型纤毛和细胞外蛋白酶而产生作用。但从腐蹄病发病症状看,节瘤拟杆菌所致疾病的严重程度不是相同的,据此节瘤拟杆菌被分为毒性、弱毒性和良性菌,这种分类通过对该菌基因组毒性关联蛋白Vap(Virulence-associated protein)区域和毒性相关位点vrl(Virulence Related Locus)区域的研究,发现其致病特点与基因的顺序有很大联系。     

拟南芥和水稻cystatin基因家族的生物信息学分析

利用已经分离的植物半胱氨酸蛋白酶抑制剂(cystatin)基因的cystatin结构域为检索序列在基因组水平上对拟南芥和水稻中的cystatin基因家族的成员进行分析;同时利用这些基因编码的蛋白质序列构建系统发生树,并对这些蛋白序列的保守序列进行分析;最后在GenBank的EST数据库中查找这些基因的ESTs表达序列。结果表明:1结合结构域的鉴定、多序列联配以及MEME分析最终确定了7个拟南芥和11个水稻的cystatin基因。2系统发生分析表明,cystatin基因的基本特征很可能是在拟南芥和水稻的分离之前就已经形成。3 cystatin结构域在蛋白质间高度保守。4拟南芥和水稻的cystatin基因主要在花、叶、根、种子和愈伤组织中表达,这有助于植物避免昆虫的侵害。