微生物学报 Pub Date : 2017-01-04

Manli Wu, Xiaoyu Li, Nan Zhang, Shuang Xu, Zhiqiang Liu

{"title":"[Gene cloning and biological function of CgRGS2 in Colletotrichum gloeosporioides].","authors":"Manli Wu, Xiaoyu Li, Nan Zhang, Shuang Xu, Zhiqiang Liu","doi":"","DOIUrl":"","url":null,"abstract":"Objective: Regulators of G-protein signaling (RGS) are negative regulatory factors of G protein and play important roles in growth development and pathogenicity of plant pathogen. However, biological functions of RGS in Colletotrichum gloeosporioides have not been studied so far. We cloned an RGS gene of CgRGS2 in C. gloeosporioides and analyzed its biological function.Methods: Gene CgRGS2 was cloned using PCR and analyzed. The gene-knockout mutant of CgRGS2 was obtained by homologous recombination, and the complementary strain was also built based on the mutant. Biological function of CgRGS2 was determined through phenotypic analysis.Results: CgRGS2 encoded a 574-amino acids protein, containing an RGS function domain in the N terminal. Comparing to the wild type, the knockout mutant of CgRGS2 had slow growth, thick aerial hyphae, reduced conidia with multi-end germination, sensitive to oxidative stress and SDS, decreased pathogenicity.Conclusion: Protein CgRGS2 was involved in regulation of vegetative growth, conidium production and germination, oxidative stress response, cell wall integrity and pathogenicity of C. gloeosporioides.","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"57 1","pages":"66-76"},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Purification and characterization of endoglucanase Egn21 from Fusarium sp. Q7-31T]. [镰刀菌Q7-31T内切葡聚糖酶Egn21的纯化及特性研究]。

微生物学报 Pub Date : 2017-01-04

Xinyuan Chang, Zhanling Xie, Fengmei Zhang, Jieqiong Lei, Rongwei Cui, Shouyi Nie

引用次数: 0

[Mutant construction and characterization of hfq in Mesorhizobium huakuii 7653R]. [华奎中干酪根菌7653R hfq突变体的构建与鉴定]。

微生物学报 Pub Date : 2017-01-04

Chuncao Ma, Xuejuan Zhou, Fuli Xie, Youguo Li

引用次数: 0

[Interaction between microflora and nitrogen nutrients in large intestine and its impacts on host health]. [大肠菌群与氮营养物质的相互作用及其对宿主健康的影响]。

微生物学报 Pub Date : 2017-01-04

Zhuang Liu, Junshi Shen, Weiyun Zhu

引用次数: 0

[The nuclear import of Newcastle disease virus matrix protein depends on KPNB1 and Ran protein]. [新城疫病毒基质蛋白的核输入取决于KPNB1和Ran蛋白]。

微生物学报 Pub Date : 2017-01-04

Zhiqiang Duan, Xinqin Ji, Jouqiang Xu, Jiafu Zhao, Haixu Xu, Shunlin Hu, Xiufan Liu

{"title":"[The nuclear import of Newcastle disease virus matrix protein depends on KPNB1 and Ran protein].","authors":"Zhiqiang Duan, Xinqin Ji, Jouqiang Xu, Jiafu Zhao, Haixu Xu, Shunlin Hu, Xiufan Liu","doi":"","DOIUrl":"","url":null,"abstract":"Objective: The aim of this study was to identify the transport proteins that mediates the nuclear import of Newcastle disease virus (NDV) matrix (M) protein.Methods: Chicken KPNA1 to KPNA6 gene and KPNB1 gene were cloned from DF-1 cells and then inserted into eukaryotic expression vectors. The constructed recombinant plasmids with a combination of grouping were transfected into HEK-293T cells to identify the transport proteins interacting with NDV M protein by co-immunoprecipitation (Co-IP) assay. Moreover, fluorescent co-localization assay was used to verify the transport proteins by co-expressing M and Ran protein mutant or M and its interactive protein deletant.Results: The recombinant proteins could normally express in plasmid-transfected HEK-293T cells. Indirect immunofluorescence detection showed that the recombinant proteins except for Myc-KPNA2 displayed the same nuclear localization as NDV M protein. The results of Co-IP revealed that M protein could interact with KPNA1 and KPNB1. Further fluorescent co-localization indicated that co-expression of M and DN-KPNA1 did not change the nuclear localization of M, whereas co-expression of M and DN-KPNB1 or M and Ran-Q69L disrupted the nuclear localization of M, demonstrating that the nuclear import of M protein was dependent on KPNB1 and Ran protein.Conclusion: KPNB1 and Ran protein jointly mediated the nuclear import of NDV M protein, showing that KPNB1 protein interacted with NDV M protein to form binary complex and then entered into the nucleus with the assistance of Ran protein.","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"57 1","pages":"109-20"},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Construction of recombinant Bacillus subtilis by co-expression of heterologous D-hydantoinase and N-carbamoylase]. [异种d -羟脲酶和n -氨基淀粉酶共表达构建重组枯草芽孢杆菌]。

微生物学报 Pub Date : 2017-01-04

Yameng Wang, Rui Ban, Lu Liu, Yu Shen

{"title":"[Construction of recombinant Bacillus subtilis by co-expression of heterologous D-hydantoinase and N-carbamoylase].","authors":"Yameng Wang, Rui Ban, Lu Liu, Yu Shen","doi":"","DOIUrl":"","url":null,"abstract":"Objective: We aimed at co-expressing heterologous D-hydantoinase and N-carbamoylase in Bacillus subtilis, and evaluating the feasibility of producing D-p-hydroxyphenylglycine by the recombinant B. subtilis whole-cell catalysis.Methods: The Paco expression cassette was combined with the coding sequence of hyd or sd1 gene as an artificial gene to express D-hydantoinase. The PAE expression cassette was combined with the coding sequence of adc gene as an artificial gene to express N-carbamoylase. The D-hydantoinase and N-carbamoylase co-expression plasmids pHCS(sd1+adc) and pHCY(hyd+adc) were constructed, using plasmid pHP13 as carrier; the co-expression plasmids pUCS(sd1+adc) was constructed, using plasmid pUB110 as carrier. The additional copy of acoR and sigL gene was integrated at chromosome. The skf and sdp gene were knocked out in B. subtilis. All recombinant strains bearing co-expression plasmid were characterized by analyzing whole-cell catalysis activity.Results: In the recombinant strains with plasmid pHCY and with pHCS, the whole-cell catalytic activity reached 0.21 U/mL and 0.31 U/mL, respectively. After the over-expression of acoR, sigL, and high-copy-number pUCS, the whole-cell catalytic activity reached 1.0 U/mL.Conclusion: Overexpression of acoR, sigL and the deletion of skf, sdp genes had significant effects on the catalysis activity of recombinant whole-cell.","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"57 1","pages":"54-65"},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36087232","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Heterologous expression, purification and characterization of phospholipase C from Bacillus cereus in Kluyveromyces lactis]. [蜡样芽孢杆菌磷脂酶C在乳酸克鲁维菌中的异源表达、纯化和特性研究]。

微生物学报 Pub Date : 2017-01-04

Chao Xiao, Liang Zhang, Yanyan Li, Yu Xin, Guoan Chen, Shengrong Yang

{"title":"[Heterologous expression, purification and characterization of phospholipase C from Bacillus cereus in Kluyveromyces lactis].","authors":"Chao Xiao, Liang Zhang, Yanyan Li, Yu Xin, Guoan Chen, Shengrong Yang","doi":"","DOIUrl":"","url":null,"abstract":"Objective: In this study, we constructed recombinant Kluyveromyces lactis strains to produce phospholipase C (PLC) of Bacillus cereus. The recombinant enzymes were purified and characterized.Methods: We cloned the PLC encoding gene bcplc of Bacillus cereus. And the amplified fragments were inserted into pKLAC1 to obtain expression plasmids. K. lactis harboring the above plasmids was cultivated to express PLC that was purified by HisTrapTM affinity chromatography and characterized.Results: PLC of B. cereus was cloned and expressed in K. lactis. The recombinant enzyme had shown activity of 19251 U/mg when using p-nitrophenyl phosphorycholine as substrate. Purified PLC exhibited optimum temperature at 80 °C and optimal pH at 9.0. The recombinant enzyme was stable below 40 °C and pH between 7.0 and 8.0. Cu2+ and Co2+ inhibited its activity whereas Zn2+, Mn2+, Ca2+ and Mg2+ stimulated its activity.Conclusion: It is the first time to express and characterize the PLC gene in K. lactis. These research results provide reference for the study of recombinant PLC.","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"57 1","pages":"87-96"},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Complete genome sequencing of polymalic acid-producing strain Aureobasidium pullulans CCTCC M2012223]. [产聚苹果酸菌株普鲁兰金黄色葡萄球菌CCTCC M2012223全基因组测序]。

微生物学报 Pub Date : 2017-01-04

Yongkang Wang, Xiaodan Song, Xiaorong Li, Sang-tian Yang, Xiang Zou

{"title":"[Complete genome sequencing of polymalic acid-producing strain Aureobasidium pullulans CCTCC M2012223].","authors":"Yongkang Wang, Xiaodan Song, Xiaorong Li, Sang-tian Yang, Xiang Zou","doi":"","DOIUrl":"","url":null,"abstract":"Objective: To explore the genome sequence of Aureobasidium pullulans CCTCC M2012223, analyze the key genes related to the biosynthesis of important metabolites, and provide genetic background for metabolic engineering.Methods: Complete genome of A. pullulans CCTCC M2012223 was sequenced by Illumina HiSeq high throughput sequencing platform. Then, fragment assembly, gene prediction, functional annotation, and GO/COG cluster were analyzed in comparison with those of other five A. pullulans varieties.Results: The complete genome sequence of A. pullulans CCTCC M2012223 was 30756831 bp with an average GC content of 47.49%, and 9452 genes were successfully predicted. Genome-wide analysis showed that A. pullulans CCTCC M2012223 had the biggest genome assembly size. Protein sequences involved in the pullulan and polymalic acid pathway were highly conservative in all of six A. pullulans varieties. Although both A. pullulans CCTCC M2012223 and A. pullulans var. melanogenum have a close affinity, some point mutation and inserts were occurred in protein sequences involved in melanin biosynthesis.Conclusion: Genome information of A. pullulans CCTCC M2012223 was annotated and genes involved in melanin, pullulan and polymalic acid pathway were compared, which would provide a theoretical basis for genetic modification of metabolic pathway in A. pullulans.","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"57 1","pages":"97-108"},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Analysis of proposals received and funded in discipline of microbiology of the National Natural Science Foundation of China in 2016]. 【2016年国家自然科学基金微生物学科立项立项分析】。

微生物学报 Pub Date : 2017-01-04

Qiang Li, Weimin Li, Hongyan Shan, Guiqing Xiao, Mingzhang Wen, Quansheng Du

引用次数: 0

[Effects of fatty acids and oxylipins on fungal growth, sporulation and aflatoxin production in Aspergillus]. 脂肪酸和氧化脂类对曲霉真菌生长、产孢和黄曲霉毒素产生的影响。

微生物学报 Pub Date : 2017-01-01

Shijuan Yan, Wenjie Huang, Chun-Ming Liu

引用次数: 0

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