腐蹄病致病因子节瘤拟杆菌蛋白酶在大肠杆菌...

Bacteroides nodosus is the essential causative agent of ovine footrot. It produces extracellular proteases which involved in pathogenesis of footrot. In this paper, we report the subcloning of Bacteroides nodosus protease, its overexpression in E. coli and its N-terminal polypeptide sequence. The subclone library was constructed in E. coli using SphI digested original clone (15 kb) and plasmid PTZ18R and screened using immunological assay. The expression was observed using SDS-PAGE. The subcloned DNA fragment was then cut with Sau3AI, cloned into pKK232-8 vector to perform promoter isolation and analysis. The promoter strength was determined using spectrophotometric assay.     

苏芸金杆菌感染粘虫后中肠组织学病变的研究

本文描述了Bacillus thuringiensis HD-1纯晶体感染粘虫Mythimna separata 5龄初幼虫后,其中肠肠壁细胞的超微结构变化.电镜观察表明:首先线粒体,发生病变,随后,细胞顶部肿胀、胞质电子密度下降,顶部胞质之下的各种细胞器均受损伤.随着病变加重,柱状细胞基膜内褶及杯状细胞质突起,这两个富含线粒体的区域发生病变最为明显.最终,肠壁细胞解体从基膜脱落.     

几个对鞘翅目昆虫有毒性的苏云金杆菌菌株

<正> 苏云金杆菌Bacillus thuringiensis Berliner是一种重要的昆虫病原芽孢杆菌。80年代以前注意力主要集中在对鳞翅目和双翅目昆虫有毒杀力的菌株上。进入80年代以后,Krieg等和Herrnstadt等先后分离出两株对鞘翅目昆虫敏感而对鳞翅目和双翅目不敏感的菌株。并把它们分别命名为B.thuringiensis var.tenebrionis和B.t.var.sandiego。由于     

一株产碱极端嗜盐杆菌

从青海省大柴旦盐湖中分离到一株极端嗜盐杆菌。该菌株在培养过程中产碱,革兰氏阴性,细胞杆状,0．7-1.0×2-3μm,极生鞭毛运动,绝对好氧,以氨基酸作唯一碳源。不利用碳水化合物,生长所需盐(NaCl)浓度在1 2％以上,最适盐浓度为l 8％。生长pH范围6-l0,最适pH9。MgSO4·7H2Oo-3％对生长无明显影响。细胞蛋白质酸性,不含二氨莲庚二酸和胞壁酸,含甘油二醚键化合物,不具有色素。根据以上特征,将该菌株定为一个新种,定名为产碱嗜盐杆菌(Halobocierium haloalcaligenum n.sp.)。     

球形芽孢杆菌Ts—1毒蛋白的分离纯化

Bacillus sphaericus strain Ts-1 is highly insecticidal to larvae of the mosquito. It's insecticidal component is toxic proteins. The toxin was extracted from spore-crystal complexes by disruption in a Sonicator Cell Disruptor Model W-220F followed by treatment with 0.05 mol/L NaOH. Fraction recovered from chromatography of the spore-crystal complexes on column of Sephadex G-200 were assayed against mosquito larvae and the toxic fractions from gel chromatography were subjected to SDS-PAGE. The toxic proteins in B. sphaericus Ts-1 spore-crystal complex migrated in position corresponding to 42kD and 43kD. Bioassay of the two purified proteins prepared by PAGE indicated that they were all toxic to mosquito larvae. Toxic protein was further purified by DEAE-cellulose chromatography. The toxic protein with a molecular weight of 42kD was obtained.     

根癌病土壤杆菌(Agrobacterium tumefaciens)Ti质粒的传递性研究及其在遗传工程中的应用

一、Ti质粒的发现 根癌病土壤杆菌是引起许多植物(据文献记载有83属300余种)冠瘿(crown gall)肿瘤的病原体。有关它的研究是从1907年Smith和Townsend提出该病的病原学开始的,40年代While和Braun等人用无菌冠瘿组织证明肿瘤的发生基于肿瘤基因的转化以后,吸引了越来越多的人去进行这种基因的分离和转化研究。特别是60年代,Schilpcroort等人用免疫学和分子杂交的技术,在无菌冠瘿细胞中分别测到了细菌的抗原和细菌Ti质粒的DNA序列以后,这方面的文章更是指数地增加。     

农杆菌介导海洋草酸青霉转化体系及聚酮合酶Pks生物学功能*

为研究南海柳珊瑚共附生草酸青霉SCSGAF0023的聚酮合酶(PKS)生物学功能,采用农杆菌介导法构建草酸青霉SCSGAF0023的Pks敲除株ΔPks,比较野生菌株及ΔPks的生长发育及环境适应性差异。以草酸青霉SCSGAF0023分生孢子为受体,p0380-hygB为双元载体,成功实现草酸青霉SCSGAF0023的遗传转化。结果表明:农杆菌浓度为OD₆₀₀=0.5,在200μmol/L 乙酰丁香酮(AS)诱导下与10⁷个/ml草酸青霉SCSGAF0023孢子于25℃共孵育时转化效率最高。基于上述转化体系,成功获得Pks敲除株ΔPks,并首次证实Pks正向调控草酸青霉SCSGAF0023产孢,但不影响其对环境的适应性。这为进一步系统研究真菌PKSs及聚酮化合物对真菌生长发育与环境适应性的影响提供素材。