Cloning and sequencing of the 3-phosphoglycerate kinase (PGK) gene from Penicillium citrinum and its application to heterologous gene expression

The gene coding for 3-phosphoglycerate kinase (PGK) in ML-236B (compactin)-producing Penicillium citrinum was isolated from the recombinant phage lambda library using the corresponding Aspergillus nidulans pgk gene as a probe. The P. citrinum pgk gene has an open reading frame of 1,254 bp, encoding a protein of 417 amino acids with a predicted molecular weight of 44,079 daltons. The position of the two introns, 59 and 60 bp respectively, was deduced from an homology comparison with the sequence of the A. nidulans pgk gene. The PGK protein of P. citrinum shows extensive high homology to the PGKs of four other fungi: P. chrysogenum (93%), A. nidulans (84%), Trichoderma reesei (78%) and Saccharomyces cerevisiae (68%). Almost total conservation is found in P. citrinum of residues thought to be important for the structure and function of the yeast enzyme. The strong codon preference found has greater similarity to that in other filamentous fungi than in yeast. A DNA fragment encompassing the pgk gene was shown to hybridize a 1.35-kb poly(A)⁺RNA, sufficient to encode the PGK polypeptide. A fused gene, pgk-hpt, containing the putative pgk promoter and the open reading frame of the Escherichia coli hygromycin B phospho-transferase (hpt) gene was constructed, and was successfully used to transform P. citrinum to a hygromycin B (HmB)-resistant phenotype.     

Nuclear DNA-binding proteins which recognize the intergenic control region of penicillin biosynthetic genes

The biosynthesis of penicillin, a secondary metabolite produced by Penicillium chrysogenum, is subject to sophisticated genetic and metabolic regulation. The structural genes, pcbC and pcbAB, which encode two of the penicillin biosynthetic enzymes are separated by a 1.16-kb intergenic region and transcribed divergently from one another. To identify and characterize nuclear proteins which interact with the pcbAB-pcbC intergenic promoter region, crude and partially purified nuclear extracts were used in mobility shift and DNA footprinting assays. Multiple DNA-binding proteins appear to bind to different regions of this DNA segment. An abundant nuclear protein, nuclear factor A (NF-A), binds at a single site in the intergenic promoter region and recognizes an 8-bp sequence, GCCAAGCC. Penicillin production is sensitive to nitrogen catabolite repression. The global-acting nitrogen regulatory protein NIT2 of Neurospora crassa binds strongly to the intergenic promoter region of the pcbAB and pcbC genes at a single site that contains two closely spaced GATA sequences.     

圆弧青霉提取物诱发人羊膜细胞程序外DNA合成及合成抑制的研究

从食管癌高发区河南省林县的粮食中,分离出5株圆弧青霉(Penicillium cyclopiumWestling)用大米做培养基,25℃培养两周,经氯仿:甲醇等连续提取后,得到棕褐色膏状提取物。用其中的4株,诱发出了人羊膜FL细胞株的程序外DNA合成(Unscheduled DNA Synthesis,UDS)与DNA合成抑制(DNA Synthesis Inhibition,DSI),与溶剂对照比较,具有显著性差异(P<0.05),并在一定程度上存在剂量效应关系;加S_9后,其生物学活性明显降低,推测此提取物中含有能引起人羊膜细胞DNA损伤的直接诱变剂。     

Recurrent hemoptysis with Penicillium marneffei and Stenotrophomonas maltophilia in Job’s syndrome

Pulmonary infection caused by the opportunistic organisms Penicillium marneffei and Stenotrophomonas maltophilia in patients with Job’s syndrome is rare and not well documented. The case of a 30-year-old man with Job’s syndrome who developed recurrent pneumonia and lung abscesses caused by P marneffei and S maltophilia, complicated by massive hemoptysis, is described. Bronchial artery embolization was successful in controlling the hemoptysis; however, the infection proved fatal despite appropriate antimicrobial therapy. A brief review of the literature on Job’s syndrome and its associated infective pulmonary manifestations is also presented.     

Intergeneric relationship between the Aspergillus ochraceous virus F and the Penicillium stoloniferum virus S

It was reported that the ‘slow’ component (PsV-S) of Penicillium stoloniferum virus complex also occurred in a second genus, Aspergillus ochraceous. The responsible virus for this intergeneric occurrence was considered to be the ‘fast’ component (AoV-F) of A. ochraceous virus complex. In this investigation, AoV dsRNA 1, that was previously shown to cross-hybridize with PsV-S dsRNA, has been cloned. It was 1754 bp in length and contained one open reading frame of 539 amino acids (p63), had the same genome organization as PsV-S dsRNA S1 and also had the conserved sequence motif of the PsV-S dsRNAs (5′-GCGCAAAA-3′) at the 5′ terminus. A BLAST search indicated that p63 was a putative dsRNA-dependent RNA polymerase (RdRp), had 81% of sequence homology to members of the genus Partitivirus, and grouped together with PsV-S in phylogenetic analysis. But immunoblot analysis showed that the capsid protein (P3) of AoV-F virus component did not reacted against PsV-S antiserum. These evidences suggest that the cross serological relationship between AoV-F and PsV-S previously observed may have been due to the RdRps of the respective viruses rather than between their respective capsid proteins as was assumed in 1985.     

小鼠播散性马尔尼菲青霉菌感染模型的建立及其生存率分析

目的探索建立播散性马尔尼菲青霉感染小鼠实验的方法及比较不同菌株及不同接种剂量的感染小鼠的生存率。方法以两株不同菌株及不同剂量的马尔尼菲青霉孢子经腹腔接种雌性BALB/C小鼠,观察接种后小鼠生存时间及生存率,同时检测不同时间小鼠肝脏菌落形成单位计数。结果不同接种剂量及不同菌株的感染小鼠有不同的生存率:在27～30d,ATCC182242.0×109cfu/mL组(100%)与ATCC182242.0×108cfu/mL组(17%)的死亡率比较差异有统计学意义(P0.01);B-63232.0×108cfu/mL组(100%)与ATCC182242.0×108cfu/mL组(17%)死亡率比较差异有统计学意义(P0.01)。结论成功建立了小鼠播散性马尔尼菲青霉感染模型;发现了同一菌株不同感染剂量小鼠有着不同的生存率,同一感染剂量不同菌株能造成小鼠不同的生存率。     

马尔尼菲青霉SUMS0152株有氧和厌氧环境下生理学特性研究

目的：研究有氧条件、厌氧条件和不同微量元素对马尔尼菲青霉SUMS0152的生长、色素产生的影响。方法：在有氧条件和厌氧条件下检测马尔尼菲青霉SUMS0152在沙氏培养基和含不同微量元素的培养基中的生长曲线,菌落及镜下形态特点。结果：在沙氏培养基中,菌丝相和酵母相的马尔尼菲青霉在厌氧条件下均比在有氧条件下生长快速,菌落及镜下形态无明显异常。无论有氧还是厌氧条件下,菌丝相和酵母相的马尔尼菲青霉均在缺钾培养基中生长最慢。缺钾和缺铁培养基中菌丝相马尔尼菲青霉可溶性色素产生部分被抑制。结论：马尔尼菲青霉SUMS0152更适应于厌氧条件下生长。微量元素钾、铁对马尔尼菲青霉SUMS0152的生长、色素产生有重要的影响。     

Environmental Allergens and Asthma Morbidity in Low-Income Children

Asthma morbidity is high in inner-city children in the United States, which may be related in part to increased allergens in poorly maintained housing. This study examined asthma morbidity in relation to mold, cockroach, dust mite, and cat allergens in the homes of 61 low-income Chicago children with asthma. Children exposed to higher levels of Penicillium in the bedroom had more frequent asthma symptoms, whereas those exposed to higher levels of cockroach allergen in the bedroom had a higher number of asthma symptoms. Respiratory infections confounded the association of cockroach allergen with number of asthma symptoms.     

(+)-Cyclopenol,a new naturally occurring 7-membered 2,5-dioxopiperazine alkaloid from the fungus Penicillium sclerotiorum endogenous with the Chinese mangrove Bruguiera gymnorrhiza

One new naturally occurring 7-membered 2,5-dioxopiperazine alkaloid named (+)-cyclopenol (1), along with nine known compounds including viridicatol (2), 3-(dimethylaminomethyl)-1-(1,1-dimethyl-2-propenyl)indole (3), anacine (4), aurantiomide C (5), viridicatin (6), 3-O-methylviridicatin (7), verrucosidin (8), ergosterol (9), and ergosterol peroxide (10), was isolated from the EtOAc extract of fungus Penicillium sclerotiorum, an endophytic fungal strain isolated from Chinese mangrove Bruguiera gymnorrhiza. The chemical structure of the new compound 1 was elucidated on the basis of detailed spectroscopic analysis. The absolute configuration of 1 was determined by single-crystal X-ray analysis with Cu Kα radiation (λ = 1.54178 Å). To our knowledge, (+)-cyclopenol (1) represents the first example of 7-membered 2,5-dioxopiperazine isolated from mangrove endophytic fungus.