In addition to ergosterol (I), 3b, 5a, 6b-trihydroxyergosta-7, 22-diene (II), 3b-hydroxy-ergosta-5-ene (Ⅲ), 3-oxo-ergosta-4, 6, 8 (14), 22-tetraene (IV), 3b-hydroxy-5a, 8a-epidioxy-ergosta-6, 22-diene (V), 3b-hydroxy-5a,8a-epidioxy-ergosta-6, 9 (11), 22-triene (VI) and 3-oxo-ergosta-4-ene (VII), a plant hormone indole-3-acetic acid (IAA) and three new antimicrobial metabolites were characterized from the culture of Colletotrichum sp., an endophyte isolated from inside the stem of Artemisia annua. The structures of the new metabolites were elucidated by a combination of spectroscopic methods (IR, MS, 1H and 13C NMR) as 6-isoprenylindole-3-carboxylic acid (1), 3b,5a-dihydroxy-6b-acetoxy-ergosta-7, 22-diene (2) and 3b, 5a-dihydroxy-6b-phenylacetyloxy-ergosta-7, 22-diene (3), respectively. The compounds 1-3 and Ⅲ-V inhibited the growth of all the tested bacteria (Bacillus subtilis, Staphylococcus aureus, Sarcina lutea and Pseudomonas sp.) with minimal inhibitory concentrations (MICs) ranging from 25 to 75 mg: ml. Moreover, metabolites 2 and 3, together with the known sterols III and V, were inhibitory against the fungi Candida albicans and Aspergillus niger with MICs between 50 and 100 mg: ml. At 200 mg: ml, compounds 1-3, III and IV were shown to be fungistatic to the crop pathogenic fungi Gaeumannomyces graminis var. tritici, Rhizoctonia cerealis, Helminthosporium sati6um and Phytophthora capisici. This is the first report on the endophytic fungus from A. annua and the bioactive metabolites thereof.

Artemisia annua, well recognized for its production of antimalarial drug artemisinin, is seldom attacked by any of phytopathogenic fungi, which could be partially associated with the presence of endophytes. Present investigation is aiming at disclosing whether the endophytes inside A. annua produce antifungal substances. A total of 39 endophytes were isolated and fermented, and the ferment broth was evaluated in vitro for the antifungal activity against crop-threatening fungi Gaeumannomyces graminis var. tritici, Rhizoctonia cerealis, Helminthosporium sati um, Fusarium graminearum, Gerlachia ni alis and Phytophthora capsici. These plant pathogens are still causing wheat take-all, sharp eyespot, common rot, scab, snow mould, and pepper phytophthora blight, respectively. Out of 39 endophytes investigated, 21 can produce in vitro substances that are inhibitory to all or a few of the tested phytopathogens whereas the rest yielded nothing active. Moreover, the most active broth of endophyte IV403 was extracted with EtOAc and n-butanol, and comparisons of the antifungal activity of the extracts indicated that the major active metabolites were EtOAc-extractable.

Bcl-2/adenovirus E1B 19 kDa interacting protein 2-like, BNIP-2-like (BNIPL) is a recently cloned and characterized apoptosis-associated protein that shares 72% homology with BNIP-2. It is highly expressed in human placenta and lung. A yeast two-hybrid system was used to obtain two BNIPL-interacting proteins, MIF (macrophage migration inhibitory factor) and GFER (growth factor erv1 (Saccharomyces cerevisiae)-like). The interactions were con

The human novel gene pp5644 (GeneBank Accession No. AF289559) coding for 124 amino acids was recently cloned. Overexpression of pp5644 in Hela cells significantly inhibited the growth and colony formation. The pp5644-interacting protein FAPP1 (phosphatidylinositol-four-phosphate adaptor protein1) associated protein-1, called FASP1, was obtained by using yeast two-hybrid system. The interaction between pp5644 and FASP1 was experimentally confirmed by GST pull-down assay in vitro and co-immunoprecipitation assay in vivo. Co-localization of pp5644 and FASP1 in cytoplasm in Hela cells could further support the interaction. Based on the experimental results, it is suggested that pp5644 physically bind to FASP1 and the biological significance of this kind of interaction in vivo is discussed.

化学合成人纤溶蛋白酶原K5（pK5）的编码基因并克隆到毕赤氏酵母表达系统的分泌型载体pPIC9K上，将重组质粒经BglⅡ单酶切后电转化Pichia pastoris GS115菌株，筛选出对G418有高抗性和在MM培养基上生长缓慢的转化子。经摇瓶发酵和甲醇诱导后，用15%SDS-PAGE检测发酵上清液，表明有重组蛋白pK5的高表达。经CM.Sepherose离子交换柱和Superdex 75分子筛层析两步分离纯化，获得了纯度达到98%的rpK5。用MTT方法检测的结果表明，纯化的rpK5可显著地抑制人血管内皮细胞的生长。