Expression of a β-Lactamase Preproinsulin Fusion Protein in Escherichia Coli
The level of expression of a β-lactamase human preproinsulin fusion protein 1 in Escherichia coli was increased by plasmid and host manipulations. The initial expression level (0.01% of total protein) was increased two- to three-fold by replacing the p-lactamase promoter with two strong promoters: t...
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| Veröffentlicht in: | Bio/Technology 1984-02, Vol.2 (2), p.165-168 |
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| creator | Emerick, Anne W Bertolani, Barbara L Ben-Bassat, A White, Thomas J Konrad, Michael W |
| description | The level of expression of a β-lactamase human preproinsulin fusion protein
1
in
Escherichia coli
was increased by plasmid and host manipulations. The initial expression level (0.01% of total protein) was increased two- to three-fold by replacing the p-lactamase promoter with two strong promoters:
trp
from
Serratia marcescens
and
lac
UV5 from
E. coli.
Replacement of the β-lactamase ribosome binding site (RBS) with a
trp
RBS enhanced the yield to a total of 0.16% of the total cell protein. Proinsulin was unstable, with a half-life of 14 minutes at 37°C. Proinsulin expression was increased three-fold (to about 0.5% of the total cell protein) by incubating the host at 30°C rather than 37°C. |
| doi_str_mv | 10.1038/nbt0284-165 |
| format | Article |
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1
in
Escherichia coli
was increased by plasmid and host manipulations. The initial expression level (0.01% of total protein) was increased two- to three-fold by replacing the p-lactamase promoter with two strong promoters:
trp
from
Serratia marcescens
and
lac
UV5 from
E. coli.
Replacement of the β-lactamase ribosome binding site (RBS) with a
trp
RBS enhanced the yield to a total of 0.16% of the total cell protein. Proinsulin was unstable, with a half-life of 14 minutes at 37°C. Proinsulin expression was increased three-fold (to about 0.5% of the total cell protein) by incubating the host at 30°C rather than 37°C.</description><identifier>ISSN: 0733-222X</identifier><identifier>ISSN: 1087-0156</identifier><identifier>EISSN: 2331-3684</identifier><identifier>EISSN: 1546-1696</identifier><identifier>DOI: 10.1038/nbt0284-165</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>Agriculture ; Bioinformatics ; Biomedical and Life Sciences ; Biomedical Engineering/Biotechnology ; Biomedicine ; Biotechnology ; Life Sciences</subject><ispartof>Bio/Technology, 1984-02, Vol.2 (2), p.165-168</ispartof><rights>Nature Publishing Company 1984</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c294t-a65e0817bb7fa637fc87ff8384b54bcfef7773a571b3a162314cc8e02cb498b73</citedby><cites>FETCH-LOGICAL-c294t-a65e0817bb7fa637fc87ff8384b54bcfef7773a571b3a162314cc8e02cb498b73</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nbt0284-165$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nbt0284-165$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,2725,27923,27924,41487,42556,51318</link.rule.ids></links><search><creatorcontrib>Emerick, Anne W</creatorcontrib><creatorcontrib>Bertolani, Barbara L</creatorcontrib><creatorcontrib>Ben-Bassat, A</creatorcontrib><creatorcontrib>White, Thomas J</creatorcontrib><creatorcontrib>Konrad, Michael W</creatorcontrib><title>Expression of a β-Lactamase Preproinsulin Fusion Protein in Escherichia Coli</title><title>Bio/Technology</title><addtitle>Nat Biotechnol</addtitle><description>The level of expression of a β-lactamase human preproinsulin fusion protein
1
in
Escherichia coli
was increased by plasmid and host manipulations. The initial expression level (0.01% of total protein) was increased two- to three-fold by replacing the p-lactamase promoter with two strong promoters:
trp
from
Serratia marcescens
and
lac
UV5 from
E. coli.
Replacement of the β-lactamase ribosome binding site (RBS) with a
trp
RBS enhanced the yield to a total of 0.16% of the total cell protein. Proinsulin was unstable, with a half-life of 14 minutes at 37°C. Proinsulin expression was increased three-fold (to about 0.5% of the total cell protein) by incubating the host at 30°C rather than 37°C.</description><subject>Agriculture</subject><subject>Bioinformatics</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedical Engineering/Biotechnology</subject><subject>Biomedicine</subject><subject>Biotechnology</subject><subject>Life Sciences</subject><issn>0733-222X</issn><issn>1087-0156</issn><issn>2331-3684</issn><issn>1546-1696</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1984</creationdate><recordtype>article</recordtype><recordid>eNp1kMFKAzEURYMoWKsrf2BcazTJy0zSpQytCiN2oeBuSEJiU9qkJDOgv-WH9JscbRE3woPLg8PlchA6p-SaEpA3QXeESY5pVR6gEQOgGCrJD9GICADMGHs9Ric5LwnhomJ8hB6n75tkc_YxFNEVqth-4kaZTq1VtsU82U2KPuR-5UMx63-weYqdHd7hptksbPJm4VVRx5U_RUdOrbI92-cYvcymz_U9bp7uHurbBhs24R1WVWmJpEJr4VQFwhkpnJMguS65Ns46IQSoUlANilYMKDdGWsKM5hOpBYzR5a7XpJhzsq7dJL9W6aOlpP020e5NtIOJgb7a0XmgwptN7TL2KQwD_8EvdnhQXZ_sb_UfufAFME1sag</recordid><startdate>19840201</startdate><enddate>19840201</enddate><creator>Emerick, Anne W</creator><creator>Bertolani, Barbara L</creator><creator>Ben-Bassat, A</creator><creator>White, Thomas J</creator><creator>Konrad, Michael W</creator><general>Nature Publishing Group US</general><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>19840201</creationdate><title>Expression of a β-Lactamase Preproinsulin Fusion Protein in Escherichia Coli</title><author>Emerick, Anne W ; Bertolani, Barbara L ; Ben-Bassat, A ; White, Thomas J ; Konrad, Michael W</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c294t-a65e0817bb7fa637fc87ff8384b54bcfef7773a571b3a162314cc8e02cb498b73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1984</creationdate><topic>Agriculture</topic><topic>Bioinformatics</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedical Engineering/Biotechnology</topic><topic>Biomedicine</topic><topic>Biotechnology</topic><topic>Life Sciences</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Emerick, Anne W</creatorcontrib><creatorcontrib>Bertolani, Barbara L</creatorcontrib><creatorcontrib>Ben-Bassat, A</creatorcontrib><creatorcontrib>White, Thomas J</creatorcontrib><creatorcontrib>Konrad, Michael W</creatorcontrib><collection>CrossRef</collection><jtitle>Bio/Technology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Emerick, Anne W</au><au>Bertolani, Barbara L</au><au>Ben-Bassat, A</au><au>White, Thomas J</au><au>Konrad, Michael W</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of a β-Lactamase Preproinsulin Fusion Protein in Escherichia Coli</atitle><jtitle>Bio/Technology</jtitle><stitle>Nat Biotechnol</stitle><date>1984-02-01</date><risdate>1984</risdate><volume>2</volume><issue>2</issue><spage>165</spage><epage>168</epage><pages>165-168</pages><issn>0733-222X</issn><issn>1087-0156</issn><eissn>2331-3684</eissn><eissn>1546-1696</eissn><abstract>The level of expression of a β-lactamase human preproinsulin fusion protein
1
in
Escherichia coli
was increased by plasmid and host manipulations. The initial expression level (0.01% of total protein) was increased two- to three-fold by replacing the p-lactamase promoter with two strong promoters:
trp
from
Serratia marcescens
and
lac
UV5 from
E. coli.
Replacement of the β-lactamase ribosome binding site (RBS) with a
trp
RBS enhanced the yield to a total of 0.16% of the total cell protein. Proinsulin was unstable, with a half-life of 14 minutes at 37°C. Proinsulin expression was increased three-fold (to about 0.5% of the total cell protein) by incubating the host at 30°C rather than 37°C.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><doi>10.1038/nbt0284-165</doi><tpages>4</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0733-222X |
| ispartof | Bio/Technology, 1984-02, Vol.2 (2), p.165-168 |
| issn | 0733-222X 1087-0156 2331-3684 1546-1696 |
| language | eng |
| recordid | cdi_crossref_primary_10_1038_nbt0284_165 |
| source | Nature Journals Online; SpringerLink Journals - AutoHoldings |
| subjects | Agriculture Bioinformatics Biomedical and Life Sciences Biomedical Engineering/Biotechnology Biomedicine Biotechnology Life Sciences |
| title | Expression of a β-Lactamase Preproinsulin Fusion Protein in Escherichia Coli |
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