progressive deletions Search Results


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  • 97
    ATCC m neoaurum atcc 25795
    ( a ) Partial gene cluster encoding the catabolism of sterols in M. <t>neoaurum</t> ATCC 25795. Genes in the map are color-coded according to assigned or proposed function: orange, mce cluster genes for steroids transportation; green, side-chain degradation genes; blue, nucleus cleavage genes; gray, unassigned function; yellow, gene encoding the transcriptional repressor KstR1. The numbers, e.g. 55 bp and 646 bp, beside the meander lines indicate the spaces between adjacent genes. ( b ) Schematic of the genomic organization of hsd4A homologues in M. neoaurum <t>ATCC</t> 25795 and other mycobacteria. Percentages, such as 71%, 81% and 67%, indicate the amino acid sequence identity of Hsd4A MN in M. neoaurum ATCC 25795 with homologs from other mycobacteria, including M. tuberculosis H37Rv, M. smegmatis str. MC 2 155 and R . jostii RHA1. The location of hsd4A is between the yrbE4B-4A and fadE26 – 27 . The size and direction of genes are indicated by arrows with corresponding length to scale.
    M Neoaurum Atcc 25795, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 211 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher pmir report vector
    ( a ) Partial gene cluster encoding the catabolism of sterols in M. <t>neoaurum</t> ATCC 25795. Genes in the map are color-coded according to assigned or proposed function: orange, mce cluster genes for steroids transportation; green, side-chain degradation genes; blue, nucleus cleavage genes; gray, unassigned function; yellow, gene encoding the transcriptional repressor KstR1. The numbers, e.g. 55 bp and 646 bp, beside the meander lines indicate the spaces between adjacent genes. ( b ) Schematic of the genomic organization of hsd4A homologues in M. neoaurum <t>ATCC</t> 25795 and other mycobacteria. Percentages, such as 71%, 81% and 67%, indicate the amino acid sequence identity of Hsd4A MN in M. neoaurum ATCC 25795 with homologs from other mycobacteria, including M. tuberculosis H37Rv, M. smegmatis str. MC 2 155 and R . jostii RHA1. The location of hsd4A is between the yrbE4B-4A and fadE26 – 27 . The size and direction of genes are indicated by arrows with corresponding length to scale.
    Pmir Report Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4287 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Illumina Inc 2bp deletion
    ( a ) Partial gene cluster encoding the catabolism of sterols in M. <t>neoaurum</t> ATCC 25795. Genes in the map are color-coded according to assigned or proposed function: orange, mce cluster genes for steroids transportation; green, side-chain degradation genes; blue, nucleus cleavage genes; gray, unassigned function; yellow, gene encoding the transcriptional repressor KstR1. The numbers, e.g. 55 bp and 646 bp, beside the meander lines indicate the spaces between adjacent genes. ( b ) Schematic of the genomic organization of hsd4A homologues in M. neoaurum <t>ATCC</t> 25795 and other mycobacteria. Percentages, such as 71%, 81% and 67%, indicate the amino acid sequence identity of Hsd4A MN in M. neoaurum ATCC 25795 with homologs from other mycobacteria, including M. tuberculosis H37Rv, M. smegmatis str. MC 2 155 and R . jostii RHA1. The location of hsd4A is between the yrbE4B-4A and fadE26 – 27 . The size and direction of genes are indicated by arrows with corresponding length to scale.
    2bp Deletion, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 85/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Angelini raven s colored progressive matrices
    ( a ) Partial gene cluster encoding the catabolism of sterols in M. <t>neoaurum</t> ATCC 25795. Genes in the map are color-coded according to assigned or proposed function: orange, mce cluster genes for steroids transportation; green, side-chain degradation genes; blue, nucleus cleavage genes; gray, unassigned function; yellow, gene encoding the transcriptional repressor KstR1. The numbers, e.g. 55 bp and 646 bp, beside the meander lines indicate the spaces between adjacent genes. ( b ) Schematic of the genomic organization of hsd4A homologues in M. neoaurum <t>ATCC</t> 25795 and other mycobacteria. Percentages, such as 71%, 81% and 67%, indicate the amino acid sequence identity of Hsd4A MN in M. neoaurum ATCC 25795 with homologs from other mycobacteria, including M. tuberculosis H37Rv, M. smegmatis str. MC 2 155 and R . jostii RHA1. The location of hsd4A is between the yrbE4B-4A and fadE26 – 27 . The size and direction of genes are indicated by arrows with corresponding length to scale.
    Raven S Colored Progressive Matrices, supplied by Angelini, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Promega erase a base kit
    ( a ) Partial gene cluster encoding the catabolism of sterols in M. <t>neoaurum</t> ATCC 25795. Genes in the map are color-coded according to assigned or proposed function: orange, mce cluster genes for steroids transportation; green, side-chain degradation genes; blue, nucleus cleavage genes; gray, unassigned function; yellow, gene encoding the transcriptional repressor KstR1. The numbers, e.g. 55 bp and 646 bp, beside the meander lines indicate the spaces between adjacent genes. ( b ) Schematic of the genomic organization of hsd4A homologues in M. neoaurum <t>ATCC</t> 25795 and other mycobacteria. Percentages, such as 71%, 81% and 67%, indicate the amino acid sequence identity of Hsd4A MN in M. neoaurum ATCC 25795 with homologs from other mycobacteria, including M. tuberculosis H37Rv, M. smegmatis str. MC 2 155 and R . jostii RHA1. The location of hsd4A is between the yrbE4B-4A and fadE26 – 27 . The size and direction of genes are indicated by arrows with corresponding length to scale.
    Erase A Base Kit, supplied by Promega, used in various techniques. Bioz Stars score: 89/100, based on 399 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    GeneDx Inc clia deletion analysis report
    ( a ) Partial gene cluster encoding the catabolism of sterols in M. <t>neoaurum</t> ATCC 25795. Genes in the map are color-coded according to assigned or proposed function: orange, mce cluster genes for steroids transportation; green, side-chain degradation genes; blue, nucleus cleavage genes; gray, unassigned function; yellow, gene encoding the transcriptional repressor KstR1. The numbers, e.g. 55 bp and 646 bp, beside the meander lines indicate the spaces between adjacent genes. ( b ) Schematic of the genomic organization of hsd4A homologues in M. neoaurum <t>ATCC</t> 25795 and other mycobacteria. Percentages, such as 71%, 81% and 67%, indicate the amino acid sequence identity of Hsd4A MN in M. neoaurum ATCC 25795 with homologs from other mycobacteria, including M. tuberculosis H37Rv, M. smegmatis str. MC 2 155 and R . jostii RHA1. The location of hsd4A is between the yrbE4B-4A and fadE26 – 27 . The size and direction of genes are indicated by arrows with corresponding length to scale.
    Clia Deletion Analysis Report, supplied by GeneDx Inc, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    TaKaRa mtdna deletions long range pcr
    ( a ) Partial gene cluster encoding the catabolism of sterols in M. <t>neoaurum</t> ATCC 25795. Genes in the map are color-coded according to assigned or proposed function: orange, mce cluster genes for steroids transportation; green, side-chain degradation genes; blue, nucleus cleavage genes; gray, unassigned function; yellow, gene encoding the transcriptional repressor KstR1. The numbers, e.g. 55 bp and 646 bp, beside the meander lines indicate the spaces between adjacent genes. ( b ) Schematic of the genomic organization of hsd4A homologues in M. neoaurum <t>ATCC</t> 25795 and other mycobacteria. Percentages, such as 71%, 81% and 67%, indicate the amino acid sequence identity of Hsd4A MN in M. neoaurum ATCC 25795 with homologs from other mycobacteria, including M. tuberculosis H37Rv, M. smegmatis str. MC 2 155 and R . jostii RHA1. The location of hsd4A is between the yrbE4B-4A and fadE26 – 27 . The size and direction of genes are indicated by arrows with corresponding length to scale.
    Mtdna Deletions Long Range Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore 6 ohda
    TH fiber density in striatum. A series of five sections from one rat is shown for each vector group. The left hemisphere is the injected side, showing marked loss of striatal TH fiber density in the <t>6-OHDA,</t> AAV8 tau, AAV9 tau or AAV10 tau vector groups.
    6 Ohda, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1832 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Promega pgl3 basic vector
    Characterization of the PURG promoter. ( A ) The PURG and WRN major transcription start sites are indicated by arrowed lines. Several potential cis -regulatory elements described in the text are indicated by open boxes. The numerical system used here is the same as in Figures 1 and 2A. ( B ) Promoter activities of various DNA fragments were measured as described in Materials and Methods. The arrows with numbers at their ends indicate the locations of various DNA fragments and the directions in which they are cloned into <t>pGL3-Basic</t> (Promega). Plasmid pGL3-Control (Promega) contains SV40 promoter and enhancer sequences that drive strong expression of the firefly luciferase gene, while plasmid pGL3-Basic lacks eukaryotic promoter and enhancer sequences. Numbers presented are relative light emission from the luciferase reaction, detected as described in Materials and Methods. Emission due to plasmid pGL3-Basic promoter activity is assigned the value 1.0.
    Pgl3 Basic Vector, supplied by Promega, used in various techniques. Bioz Stars score: 95/100, based on 27770 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Agilent technologies array based comparative genomic hybridization
    Characterization of the PURG promoter. ( A ) The PURG and WRN major transcription start sites are indicated by arrowed lines. Several potential cis -regulatory elements described in the text are indicated by open boxes. The numerical system used here is the same as in Figures 1 and 2A. ( B ) Promoter activities of various DNA fragments were measured as described in Materials and Methods. The arrows with numbers at their ends indicate the locations of various DNA fragments and the directions in which they are cloned into <t>pGL3-Basic</t> (Promega). Plasmid pGL3-Control (Promega) contains SV40 promoter and enhancer sequences that drive strong expression of the firefly luciferase gene, while plasmid pGL3-Basic lacks eukaryotic promoter and enhancer sequences. Numbers presented are relative light emission from the luciferase reaction, detected as described in Materials and Methods. Emission due to plasmid pGL3-Basic promoter activity is assigned the value 1.0.
    Array Based Comparative Genomic Hybridization, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Novartis i deleu reports clinical trial funding
    Characterization of the PURG promoter. ( A ) The PURG and WRN major transcription start sites are indicated by arrowed lines. Several potential cis -regulatory elements described in the text are indicated by open boxes. The numerical system used here is the same as in Figures 1 and 2A. ( B ) Promoter activities of various DNA fragments were measured as described in Materials and Methods. The arrows with numbers at their ends indicate the locations of various DNA fragments and the directions in which they are cloned into <t>pGL3-Basic</t> (Promega). Plasmid pGL3-Control (Promega) contains SV40 promoter and enhancer sequences that drive strong expression of the firefly luciferase gene, while plasmid pGL3-Basic lacks eukaryotic promoter and enhancer sequences. Numbers presented are relative light emission from the luciferase reaction, detected as described in Materials and Methods. Emission due to plasmid pGL3-Basic promoter activity is assigned the value 1.0.
    I Deleu Reports Clinical Trial Funding, supplied by Novartis, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    The Jackson Laboratory flp deleter mouse
    Characterization of the PURG promoter. ( A ) The PURG and WRN major transcription start sites are indicated by arrowed lines. Several potential cis -regulatory elements described in the text are indicated by open boxes. The numerical system used here is the same as in Figures 1 and 2A. ( B ) Promoter activities of various DNA fragments were measured as described in Materials and Methods. The arrows with numbers at their ends indicate the locations of various DNA fragments and the directions in which they are cloned into <t>pGL3-Basic</t> (Promega). Plasmid pGL3-Control (Promega) contains SV40 promoter and enhancer sequences that drive strong expression of the firefly luciferase gene, while plasmid pGL3-Basic lacks eukaryotic promoter and enhancer sequences. Numbers presented are relative light emission from the luciferase reaction, detected as described in Materials and Methods. Emission due to plasmid pGL3-Basic promoter activity is assigned the value 1.0.
    Flp Deleter Mouse, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 85/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Agilent technologies qc report
    Characterization of the PURG promoter. ( A ) The PURG and WRN major transcription start sites are indicated by arrowed lines. Several potential cis -regulatory elements described in the text are indicated by open boxes. The numerical system used here is the same as in Figures 1 and 2A. ( B ) Promoter activities of various DNA fragments were measured as described in Materials and Methods. The arrows with numbers at their ends indicate the locations of various DNA fragments and the directions in which they are cloned into <t>pGL3-Basic</t> (Promega). Plasmid pGL3-Control (Promega) contains SV40 promoter and enhancer sequences that drive strong expression of the firefly luciferase gene, while plasmid pGL3-Basic lacks eukaryotic promoter and enhancer sequences. Numbers presented are relative light emission from the luciferase reaction, detected as described in Materials and Methods. Emission due to plasmid pGL3-Basic promoter activity is assigned the value 1.0.
    Qc Report, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ( a ) Partial gene cluster encoding the catabolism of sterols in M. neoaurum ATCC 25795. Genes in the map are color-coded according to assigned or proposed function: orange, mce cluster genes for steroids transportation; green, side-chain degradation genes; blue, nucleus cleavage genes; gray, unassigned function; yellow, gene encoding the transcriptional repressor KstR1. The numbers, e.g. 55 bp and 646 bp, beside the meander lines indicate the spaces between adjacent genes. ( b ) Schematic of the genomic organization of hsd4A homologues in M. neoaurum ATCC 25795 and other mycobacteria. Percentages, such as 71%, 81% and 67%, indicate the amino acid sequence identity of Hsd4A MN in M. neoaurum ATCC 25795 with homologs from other mycobacteria, including M. tuberculosis H37Rv, M. smegmatis str. MC 2 155 and R . jostii RHA1. The location of hsd4A is between the yrbE4B-4A and fadE26 – 27 . The size and direction of genes are indicated by arrows with corresponding length to scale.

    Journal: Scientific Reports

    Article Title: Unraveling and engineering the production of 23,24-bisnorcholenic steroids in sterol metabolism

    doi: 10.1038/srep21928

    Figure Lengend Snippet: ( a ) Partial gene cluster encoding the catabolism of sterols in M. neoaurum ATCC 25795. Genes in the map are color-coded according to assigned or proposed function: orange, mce cluster genes for steroids transportation; green, side-chain degradation genes; blue, nucleus cleavage genes; gray, unassigned function; yellow, gene encoding the transcriptional repressor KstR1. The numbers, e.g. 55 bp and 646 bp, beside the meander lines indicate the spaces between adjacent genes. ( b ) Schematic of the genomic organization of hsd4A homologues in M. neoaurum ATCC 25795 and other mycobacteria. Percentages, such as 71%, 81% and 67%, indicate the amino acid sequence identity of Hsd4A MN in M. neoaurum ATCC 25795 with homologs from other mycobacteria, including M. tuberculosis H37Rv, M. smegmatis str. MC 2 155 and R . jostii RHA1. The location of hsd4A is between the yrbE4B-4A and fadE26 – 27 . The size and direction of genes are indicated by arrows with corresponding length to scale.

    Article Snippet: According to the steroid nucleus metabolic mechanism , kstD1 , kstD2 , kstD3 and hsd4A MN were progressively deleted in M. neoaurum ATCC 25795, resulting in the final strain MNΔhsd4A ΔkstD123 .

    Techniques: Sequencing

    Phenotypic analyses of the metabolism of sterol by M. neoaurum ATCC25795 and its derivative strains. ( a ) HPLC chromatogram comparison of the products from the transformation of 2 g/l of cholesterol in MYC/02 media at 30 °C by strains M. neoaurum ATCC 25795 (blue), NwIB-XII (red) and XIIp261 hsd4A (green). Cholesterol can be completely degraded by M. neoaurum ATCC 25795 without obvious accumulation of intermediates. The catabolism of cholesterol in NwIB-XII was blocked to accumulate multiple metabolites, including ADD (I), AD (II), TS (III) and BD (IV). The plasmid pMV261- hsd4A was electro-introduced into NwIB-XII, resulting in the XIIp261 hsd4A strain. Strain XIIp261 hsd4A transformed cholesterol to ADD (I) and AD (II). ( b ) Conversion relationship between the metabolites (I to IV). Arrows in purple signify the function of Hsd4A MN deduced from the product phenotypes of strains NwIB-XII and XIIp261 hsd4A . Hsd4A MN can irreversibly catalyze the oxidation of TS to AD and BD to ADD, respectively. AD, androst-4-ene-3,17-dione; ADD, androst-1,4-dien-3,17-dione; BD, boldenone; TS, testosterone.

    Journal: Scientific Reports

    Article Title: Unraveling and engineering the production of 23,24-bisnorcholenic steroids in sterol metabolism

    doi: 10.1038/srep21928

    Figure Lengend Snippet: Phenotypic analyses of the metabolism of sterol by M. neoaurum ATCC25795 and its derivative strains. ( a ) HPLC chromatogram comparison of the products from the transformation of 2 g/l of cholesterol in MYC/02 media at 30 °C by strains M. neoaurum ATCC 25795 (blue), NwIB-XII (red) and XIIp261 hsd4A (green). Cholesterol can be completely degraded by M. neoaurum ATCC 25795 without obvious accumulation of intermediates. The catabolism of cholesterol in NwIB-XII was blocked to accumulate multiple metabolites, including ADD (I), AD (II), TS (III) and BD (IV). The plasmid pMV261- hsd4A was electro-introduced into NwIB-XII, resulting in the XIIp261 hsd4A strain. Strain XIIp261 hsd4A transformed cholesterol to ADD (I) and AD (II). ( b ) Conversion relationship between the metabolites (I to IV). Arrows in purple signify the function of Hsd4A MN deduced from the product phenotypes of strains NwIB-XII and XIIp261 hsd4A . Hsd4A MN can irreversibly catalyze the oxidation of TS to AD and BD to ADD, respectively. AD, androst-4-ene-3,17-dione; ADD, androst-1,4-dien-3,17-dione; BD, boldenone; TS, testosterone.

    Article Snippet: According to the steroid nucleus metabolic mechanism , kstD1 , kstD2 , kstD3 and hsd4A MN were progressively deleted in M. neoaurum ATCC 25795, resulting in the final strain MNΔhsd4A ΔkstD123 .

    Techniques: High Performance Liquid Chromatography, Transformation Assay, Plasmid Preparation

    Schematic diagram of sterol catabolism and the combinatorial construction of HBCs producing strains. ( a ) The construction of strain XIIΔ hsd4A -p261 kstD1 . To construct XIIΔ hsd4A -p261 kstD1 , hsd4A was deleted in a kshAs -null mutant and then kstD1 was overexpressed. The resulting strain XIIΔ hsd4A -p261 kstD1 transformed phytosterols to 1,4-HBC as the major product. ( b ) The construction of strain XIIΔ hsd4A Δ kstD123 . To construct XIIΔ hsd4A Δ kstD123 , hsd4A was deleted in a kshAs -null mutant and then kstD1 , kstD2 and kstD3 were knocked out sequentially. The resulting strain XIIΔ hsd4A Δ kstD123 transformed phytosterols to 4-HBC as the major product. ( c ) The construction of strain MNΔ hsd4A Δ kstD123 . To construct MNΔ hsd4A Δ kstD123 , hsd4A was deleted in M. neoaurum ATCC 25795 and then kstD1 , kstD2 and kstD3 were knocked out sequentially. The resulting strain MNΔ hsd4A Δ kstD123 transformed phytosterols to 9-OHHBC as the major product. 9-OHHBC, 9,22-dihydroxy-23,24-bisnorchol-4-ene-3-one. ( d ) Proposed metabolism of the sterol side chain for the production of C19 and C22 steroids in M. neoaurum ATCC 25795. The conversion from 22HOBNC-CoA to AD was designated as the AD pathway (orange arrows) and has been proposed as the sole pathway of sterol side-chain degradation. Here, a new pathway is proposed: the conversion from 22HOBNC-CoA to 4-HBC (designated as the HBC pathway, blue arrows). Between the two pathways, 22HOBNC-CoA is the branching-node, which leads to AD via the catalysis of Hsd4A MN and leads to 4-HBC via an aldolytic reaction. The β-hydroxybutyryl-CoA moiety of 22HOBNC-CoA and acetoacetyl-CoA moiety of 22OBNC-CoA are labelled in red, which were the active substrates of Hsd4A MN in vitro . 24HOC-CoA, 24-hydroxy-3-oxo-chol-4-en-26-oyl CoA; 22HOBNC-CoA, 22-hydroxy-3-oxo-25,26-bisnorchol-4-en-24-oyl CoA; 22OBNC-CoA, 3,22-dioxo-25,26-bisnorchol-4-ene-24-oyl CoA; OBNC22-CoA, 3-oxo-22,23-bisnorchol-4-ene-22-oyl-CoA; OBNC20FA, 3-oxo-23,24-bisnorchol-4-ene-20-formic acid; OBNC20CA, 3-oxo-23,24-bisnorchol-4-ene-20-carbaldehyde. ( e ) The proposed pathway of cholate side-chain degradation in Pseudomonas sp. strain Chol1 37 . sal , an aldol-lyase; sad , an aldehyde dehydrogenase. Intercepted arrows (red) indicate the blockage of the target reaction by gene deletion. Arrows in purple signify the enhanced KstD1 activity. Compounds in green are the major products accumulated in constructed strains. The compound in brackets is an unstable compound that will lead to the decomposition of steroid nucleus and further degradation. Dashed arrows indicate the proposed way for HBCs formation previously suggested by Szentirmai 32 . Arrows in arrays signify multiple reaction steps.

    Journal: Scientific Reports

    Article Title: Unraveling and engineering the production of 23,24-bisnorcholenic steroids in sterol metabolism

    doi: 10.1038/srep21928

    Figure Lengend Snippet: Schematic diagram of sterol catabolism and the combinatorial construction of HBCs producing strains. ( a ) The construction of strain XIIΔ hsd4A -p261 kstD1 . To construct XIIΔ hsd4A -p261 kstD1 , hsd4A was deleted in a kshAs -null mutant and then kstD1 was overexpressed. The resulting strain XIIΔ hsd4A -p261 kstD1 transformed phytosterols to 1,4-HBC as the major product. ( b ) The construction of strain XIIΔ hsd4A Δ kstD123 . To construct XIIΔ hsd4A Δ kstD123 , hsd4A was deleted in a kshAs -null mutant and then kstD1 , kstD2 and kstD3 were knocked out sequentially. The resulting strain XIIΔ hsd4A Δ kstD123 transformed phytosterols to 4-HBC as the major product. ( c ) The construction of strain MNΔ hsd4A Δ kstD123 . To construct MNΔ hsd4A Δ kstD123 , hsd4A was deleted in M. neoaurum ATCC 25795 and then kstD1 , kstD2 and kstD3 were knocked out sequentially. The resulting strain MNΔ hsd4A Δ kstD123 transformed phytosterols to 9-OHHBC as the major product. 9-OHHBC, 9,22-dihydroxy-23,24-bisnorchol-4-ene-3-one. ( d ) Proposed metabolism of the sterol side chain for the production of C19 and C22 steroids in M. neoaurum ATCC 25795. The conversion from 22HOBNC-CoA to AD was designated as the AD pathway (orange arrows) and has been proposed as the sole pathway of sterol side-chain degradation. Here, a new pathway is proposed: the conversion from 22HOBNC-CoA to 4-HBC (designated as the HBC pathway, blue arrows). Between the two pathways, 22HOBNC-CoA is the branching-node, which leads to AD via the catalysis of Hsd4A MN and leads to 4-HBC via an aldolytic reaction. The β-hydroxybutyryl-CoA moiety of 22HOBNC-CoA and acetoacetyl-CoA moiety of 22OBNC-CoA are labelled in red, which were the active substrates of Hsd4A MN in vitro . 24HOC-CoA, 24-hydroxy-3-oxo-chol-4-en-26-oyl CoA; 22HOBNC-CoA, 22-hydroxy-3-oxo-25,26-bisnorchol-4-en-24-oyl CoA; 22OBNC-CoA, 3,22-dioxo-25,26-bisnorchol-4-ene-24-oyl CoA; OBNC22-CoA, 3-oxo-22,23-bisnorchol-4-ene-22-oyl-CoA; OBNC20FA, 3-oxo-23,24-bisnorchol-4-ene-20-formic acid; OBNC20CA, 3-oxo-23,24-bisnorchol-4-ene-20-carbaldehyde. ( e ) The proposed pathway of cholate side-chain degradation in Pseudomonas sp. strain Chol1 37 . sal , an aldol-lyase; sad , an aldehyde dehydrogenase. Intercepted arrows (red) indicate the blockage of the target reaction by gene deletion. Arrows in purple signify the enhanced KstD1 activity. Compounds in green are the major products accumulated in constructed strains. The compound in brackets is an unstable compound that will lead to the decomposition of steroid nucleus and further degradation. Dashed arrows indicate the proposed way for HBCs formation previously suggested by Szentirmai 32 . Arrows in arrays signify multiple reaction steps.

    Article Snippet: According to the steroid nucleus metabolic mechanism , kstD1 , kstD2 , kstD3 and hsd4A MN were progressively deleted in M. neoaurum ATCC 25795, resulting in the final strain MNΔhsd4A ΔkstD123 .

    Techniques: Construct, Mutagenesis, Transformation Assay, In Vitro, Activity Assay

    TH fiber density in striatum. A series of five sections from one rat is shown for each vector group. The left hemisphere is the injected side, showing marked loss of striatal TH fiber density in the 6-OHDA, AAV8 tau, AAV9 tau or AAV10 tau vector groups.

    Journal: The European journal of neuroscience

    Article Title: Tau expression levels from various adeno-associated virus vector serotypes produce graded neurodegenerative disease states

    doi: 10.1111/j.1460-9568.2008.06161.x

    Figure Lengend Snippet: TH fiber density in striatum. A series of five sections from one rat is shown for each vector group. The left hemisphere is the injected side, showing marked loss of striatal TH fiber density in the 6-OHDA, AAV8 tau, AAV9 tau or AAV10 tau vector groups.

    Article Snippet: Some rats were injected with 6-OHDA (Sigma, St Louis, MO, USA).

    Techniques: Plasmid Preparation, Injection

    Characterization of the PURG promoter. ( A ) The PURG and WRN major transcription start sites are indicated by arrowed lines. Several potential cis -regulatory elements described in the text are indicated by open boxes. The numerical system used here is the same as in Figures 1 and 2A. ( B ) Promoter activities of various DNA fragments were measured as described in Materials and Methods. The arrows with numbers at their ends indicate the locations of various DNA fragments and the directions in which they are cloned into pGL3-Basic (Promega). Plasmid pGL3-Control (Promega) contains SV40 promoter and enhancer sequences that drive strong expression of the firefly luciferase gene, while plasmid pGL3-Basic lacks eukaryotic promoter and enhancer sequences. Numbers presented are relative light emission from the luciferase reaction, detected as described in Materials and Methods. Emission due to plasmid pGL3-Basic promoter activity is assigned the value 1.0.

    Journal: Nucleic Acids Research

    Article Title: Distinct proteins encoded by alternative transcripts of the PURG gene, located contrapodal to WRN on chromosome 8, determined by differential termination/polyadenylation

    doi:

    Figure Lengend Snippet: Characterization of the PURG promoter. ( A ) The PURG and WRN major transcription start sites are indicated by arrowed lines. Several potential cis -regulatory elements described in the text are indicated by open boxes. The numerical system used here is the same as in Figures 1 and 2A. ( B ) Promoter activities of various DNA fragments were measured as described in Materials and Methods. The arrows with numbers at their ends indicate the locations of various DNA fragments and the directions in which they are cloned into pGL3-Basic (Promega). Plasmid pGL3-Control (Promega) contains SV40 promoter and enhancer sequences that drive strong expression of the firefly luciferase gene, while plasmid pGL3-Basic lacks eukaryotic promoter and enhancer sequences. Numbers presented are relative light emission from the luciferase reaction, detected as described in Materials and Methods. Emission due to plasmid pGL3-Basic promoter activity is assigned the value 1.0.

    Article Snippet: A series of luciferase reporter constructs were generated by inserting PURG upstream sequences with progressively deleted 5′-ends into the pGL3-Basic vector (Promega).

    Techniques: Clone Assay, Plasmid Preparation, Expressing, Luciferase, Activity Assay