Hyperproduction of aspartase of Escherichia coli K-12 by the use of a runaway plasmid vector
Plasmid pYT125 bearing the aspartase gene ( aspA) of Escherichia coli K-12 was constructed by the use of pSY343, runaway-replication plasmid vector. One of the transformants, strain MM294 (pYT125)-S maintained pYT125 stably at 37°C in a non-selective medium. This strain produced approx. 60-fold more...
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| Veröffentlicht in: | Journal of biotechnology 1987, Vol.6 (1), p.31-40 |
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| creator | Nishimura, Noriyuki Komatsubara, Saburo Taniguchi, Tomoyasu Kisumi, Masahiko |
| description | Plasmid pYT125 bearing the aspartase gene (
aspA) of
Escherichia coli K-12 was constructed by the use of pSY343, runaway-replication plasmid vector. One of the transformants, strain MM294 (pYT125)-S maintained pYT125 stably at 37°C in a non-selective medium. This strain produced approx. 60-fold more aspartase than did the control strain after 30 cell generations. Polypeptide of the
aspA product amounted to 25–30% of the total cellular protein. Plasmid DNA isolated from strain MM294 (pYT125)-S was very similar to that from unstable strain MM294 (pYT125) in length and restriction sites. The copy number was 83 in strain MM294 (pYT125)-S and was 380 in strain MM294 (pYT125). Vector pSY343 was also stabilized in cured strains, isolated from MM294 (pYT125)-S, accompanied by reduction of the copy number. Accordingly, stabilization of pYT125 was considered to be closely related to decrease of the copy number, owing to genetic alteration of MM294-S. |
| doi_str_mv | 10.1016/0168-1656(87)90043-5 |
| format | Article |
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aspA) of
Escherichia coli K-12 was constructed by the use of pSY343, runaway-replication plasmid vector. One of the transformants, strain MM294 (pYT125)-S maintained pYT125 stably at 37°C in a non-selective medium. This strain produced approx. 60-fold more aspartase than did the control strain after 30 cell generations. Polypeptide of the
aspA product amounted to 25–30% of the total cellular protein. Plasmid DNA isolated from strain MM294 (pYT125)-S was very similar to that from unstable strain MM294 (pYT125) in length and restriction sites. The copy number was 83 in strain MM294 (pYT125)-S and was 380 in strain MM294 (pYT125). Vector pSY343 was also stabilized in cured strains, isolated from MM294 (pYT125)-S, accompanied by reduction of the copy number. Accordingly, stabilization of pYT125 was considered to be closely related to decrease of the copy number, owing to genetic alteration of MM294-S.</description><identifier>ISSN: 0168-1656</identifier><identifier>EISSN: 1873-4863</identifier><identifier>DOI: 10.1016/0168-1656(87)90043-5</identifier><language>eng</language><publisher>Elsevier B.V</publisher><subject>Aspartase ; aspartate ammonia-lyase ; Escherichia coli ; genes ; Plasmid stability ; plasmids ; Runaway-replication-plasmid</subject><ispartof>Journal of biotechnology, 1987, Vol.6 (1), p.31-40</ispartof><rights>1987</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c439t-334afa8daab7f4cc15b215fd222e12f2a6ad97546b8ebe24970bf53d451778d33</citedby><cites>FETCH-LOGICAL-c439t-334afa8daab7f4cc15b215fd222e12f2a6ad97546b8ebe24970bf53d451778d33</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/0168165687900435$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,4010,27900,27901,27902,65534</link.rule.ids></links><search><creatorcontrib>Nishimura, Noriyuki</creatorcontrib><creatorcontrib>Komatsubara, Saburo</creatorcontrib><creatorcontrib>Taniguchi, Tomoyasu</creatorcontrib><creatorcontrib>Kisumi, Masahiko</creatorcontrib><title>Hyperproduction of aspartase of Escherichia coli K-12 by the use of a runaway plasmid vector</title><title>Journal of biotechnology</title><description>Plasmid pYT125 bearing the aspartase gene (
aspA) of
Escherichia coli K-12 was constructed by the use of pSY343, runaway-replication plasmid vector. One of the transformants, strain MM294 (pYT125)-S maintained pYT125 stably at 37°C in a non-selective medium. This strain produced approx. 60-fold more aspartase than did the control strain after 30 cell generations. Polypeptide of the
aspA product amounted to 25–30% of the total cellular protein. Plasmid DNA isolated from strain MM294 (pYT125)-S was very similar to that from unstable strain MM294 (pYT125) in length and restriction sites. The copy number was 83 in strain MM294 (pYT125)-S and was 380 in strain MM294 (pYT125). Vector pSY343 was also stabilized in cured strains, isolated from MM294 (pYT125)-S, accompanied by reduction of the copy number. Accordingly, stabilization of pYT125 was considered to be closely related to decrease of the copy number, owing to genetic alteration of MM294-S.</description><subject>Aspartase</subject><subject>aspartate ammonia-lyase</subject><subject>Escherichia coli</subject><subject>genes</subject><subject>Plasmid stability</subject><subject>plasmids</subject><subject>Runaway-replication-plasmid</subject><issn>0168-1656</issn><issn>1873-4863</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1987</creationdate><recordtype>article</recordtype><recordid>eNp9kE1LxDAQhoMouK7-Aw85iR6qzVebXgSR1RUXvOhNCNNkyka6bU3alf33tq549DAMA8878D6EnLP0mqUsuxlHJyxT2aXOr4o0lSJRB2TGdC4SqTNxSGZ_yDE5ifEjHaFCsRl5X-46DF1o3WB73za0rSjEDkIPEadjEe0ag7drD9S2tafPCeO03NF-jXTYM0DD0MAX7GhXQ9x4R7do-zackqMK6ohnv3tO3h4Wr_fLZPXy-HR_t0qsFEWfCCGhAu0AyryS1jJVcqYqxzlHxisOGbgiVzIrNZbIZZGnZaWEk4rluXZCzMnF_u_Y43PA2JuNjxbrGhpsh2iY1Dxjko2g3IM2tDEGrEwX_AbCzrDUTCrN5MlMnozOzY9Ko8bY7T6GY4mtx2Ci9dhYdD6MRY1r_f8PvgEdcXrp</recordid><startdate>1987</startdate><enddate>1987</enddate><creator>Nishimura, Noriyuki</creator><creator>Komatsubara, Saburo</creator><creator>Taniguchi, Tomoyasu</creator><creator>Kisumi, Masahiko</creator><general>Elsevier B.V</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>1987</creationdate><title>Hyperproduction of aspartase of Escherichia coli K-12 by the use of a runaway plasmid vector</title><author>Nishimura, Noriyuki ; Komatsubara, Saburo ; Taniguchi, Tomoyasu ; Kisumi, Masahiko</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c439t-334afa8daab7f4cc15b215fd222e12f2a6ad97546b8ebe24970bf53d451778d33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1987</creationdate><topic>Aspartase</topic><topic>aspartate ammonia-lyase</topic><topic>Escherichia coli</topic><topic>genes</topic><topic>Plasmid stability</topic><topic>plasmids</topic><topic>Runaway-replication-plasmid</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nishimura, Noriyuki</creatorcontrib><creatorcontrib>Komatsubara, Saburo</creatorcontrib><creatorcontrib>Taniguchi, Tomoyasu</creatorcontrib><creatorcontrib>Kisumi, Masahiko</creatorcontrib><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Journal of biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nishimura, Noriyuki</au><au>Komatsubara, Saburo</au><au>Taniguchi, Tomoyasu</au><au>Kisumi, Masahiko</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Hyperproduction of aspartase of Escherichia coli K-12 by the use of a runaway plasmid vector</atitle><jtitle>Journal of biotechnology</jtitle><date>1987</date><risdate>1987</risdate><volume>6</volume><issue>1</issue><spage>31</spage><epage>40</epage><pages>31-40</pages><issn>0168-1656</issn><eissn>1873-4863</eissn><abstract>Plasmid pYT125 bearing the aspartase gene (
aspA) of
Escherichia coli K-12 was constructed by the use of pSY343, runaway-replication plasmid vector. One of the transformants, strain MM294 (pYT125)-S maintained pYT125 stably at 37°C in a non-selective medium. This strain produced approx. 60-fold more aspartase than did the control strain after 30 cell generations. Polypeptide of the
aspA product amounted to 25–30% of the total cellular protein. Plasmid DNA isolated from strain MM294 (pYT125)-S was very similar to that from unstable strain MM294 (pYT125) in length and restriction sites. The copy number was 83 in strain MM294 (pYT125)-S and was 380 in strain MM294 (pYT125). Vector pSY343 was also stabilized in cured strains, isolated from MM294 (pYT125)-S, accompanied by reduction of the copy number. Accordingly, stabilization of pYT125 was considered to be closely related to decrease of the copy number, owing to genetic alteration of MM294-S.</abstract><pub>Elsevier B.V</pub><doi>10.1016/0168-1656(87)90043-5</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0168-1656 |
| ispartof | Journal of biotechnology, 1987, Vol.6 (1), p.31-40 |
| issn | 0168-1656 1873-4863 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_14826141 |
| source | Elsevier ScienceDirect Journals Complete |
| subjects | Aspartase aspartate ammonia-lyase Escherichia coli genes Plasmid stability plasmids Runaway-replication-plasmid |
| title | Hyperproduction of aspartase of Escherichia coli K-12 by the use of a runaway plasmid vector |
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