Purification of Functional Recombinant YY1 Expressed in Sf9 Insect Cells
We expressed a gene for YY1, the multifunctional mammalian transcription factor, with a recombinant baculovirus in insect cells and obtained high levels of protein in cell. lysates. A simple and efficient purification method was developed. In the final step, we used DNA affinity chromatography with...
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| Veröffentlicht in: | Protein expression and purification 1995-04, Vol.6 (2), p.169-175 |
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| container_title | Protein expression and purification |
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| creator | Momoeda, M. Kawase, M. Young, N.S. Kajigaya, S. |
| description | We expressed a gene for YY1, the multifunctional mammalian transcription factor, with a recombinant baculovirus in insect cells and obtained high levels of protein in cell. lysates. A simple and efficient purification method was developed. In the final step, we used DNA affinity chromatography with concatermerized sequence derived from the upstream region of the B19 parvovirus P6 promoter, which has strong affinity for YY1, Approximately 1.8 mg of highly purified YY1 was obtained from 10
9 Sf9 cells. Highly purified YY1 behaved as authentic YY1 on electrophoretic mobility shift assays and in enhancing transcription from the P6 promoter. YY1 produced in a baculovirus system should prove useful for
in vitro transcription assays of YY1 and related regulatory proteins. |
| doi_str_mv | 10.1006/prep.1995.1021 |
| format | Article |
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9 Sf9 cells. Highly purified YY1 behaved as authentic YY1 on electrophoretic mobility shift assays and in enhancing transcription from the P6 promoter. YY1 produced in a baculovirus system should prove useful for
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9 Sf9 cells. Highly purified YY1 behaved as authentic YY1 on electrophoretic mobility shift assays and in enhancing transcription from the P6 promoter. YY1 produced in a baculovirus system should prove useful for
in vitro transcription assays of YY1 and related regulatory proteins.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Cell Line</subject><subject>Chromatography, Affinity</subject><subject>Chromatography, Ion Exchange</subject><subject>DNA, Viral - metabolism</subject><subject>DNA-Binding Proteins - biosynthesis</subject><subject>DNA-Binding Proteins - genetics</subject><subject>DNA-Binding Proteins - isolation & purification</subject><subject>Erythroid-Specific DNA-Binding Factors</subject><subject>Gene Expression</subject><subject>Humans</subject><subject>Molecular Sequence Data</subject><subject>Nucleopolyhedrovirus</subject><subject>Parvovirus B19, Human - genetics</subject><subject>Promoter Regions, Genetic</subject><subject>Recombinant Fusion Proteins - biosynthesis</subject><subject>Recombinant Fusion Proteins - isolation & purification</subject><subject>Spodoptera</subject><subject>Transcription Factors - biosynthesis</subject><subject>Transcription Factors - genetics</subject><subject>Transcription Factors - isolation & purification</subject><subject>YY1 Transcription Factor</subject><issn>1046-5928</issn><issn>1096-0279</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kE1PwzAMhiMEGjC4ckPKiVtH0jZJc0TTxiZNAvFx2ClKU0cK6hdJi-Dfk2oTN0625dev7QehG0oWlBB-33voF1RKFsuUnqALSiRPSCrk6ZTnPGEyLc7RZQgfhFDKCZuhmeCEU84u0OZ59M46owfXtbizeD22Zsp1jV_AdE3pWt0OeL-nePUdl4UAFXYtfrUSb9sAZsBLqOtwhc6srgNcH-Mcva9Xb8tNsnt63C4fdonJMjkkvKhyVqa84sRklgG1eVGWNi00MABdFkbmGTMSYjBasNzkaSoqk2dWCGtkNkd3B9_ed58jhEE1Lph4gW6hG4MSIis4SbMoXByExncheLCq967R_kdRoiZ0akKnJnRqQhcHbo_OY9lA9Sc_sor94tCH-N6XA6-CcdAaqJyPGFTVuf-sfwH9dn0z</recordid><startdate>19950401</startdate><enddate>19950401</enddate><creator>Momoeda, M.</creator><creator>Kawase, M.</creator><creator>Young, N.S.</creator><creator>Kajigaya, S.</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19950401</creationdate><title>Purification of Functional Recombinant YY1 Expressed in Sf9 Insect Cells</title><author>Momoeda, M. ; Kawase, M. ; Young, N.S. ; Kajigaya, S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c339t-68d45b26d60c3f5e1f48bbf28ae5eeab8c9435c9e943ca754c4227dc43f77fc93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>Cell Line</topic><topic>Chromatography, Affinity</topic><topic>Chromatography, Ion Exchange</topic><topic>DNA, Viral - metabolism</topic><topic>DNA-Binding Proteins - biosynthesis</topic><topic>DNA-Binding Proteins - genetics</topic><topic>DNA-Binding Proteins - isolation & purification</topic><topic>Erythroid-Specific DNA-Binding Factors</topic><topic>Gene Expression</topic><topic>Humans</topic><topic>Molecular Sequence Data</topic><topic>Nucleopolyhedrovirus</topic><topic>Parvovirus B19, Human - genetics</topic><topic>Promoter Regions, Genetic</topic><topic>Recombinant Fusion Proteins - biosynthesis</topic><topic>Recombinant Fusion Proteins - isolation & purification</topic><topic>Spodoptera</topic><topic>Transcription Factors - biosynthesis</topic><topic>Transcription Factors - genetics</topic><topic>Transcription Factors - isolation & purification</topic><topic>YY1 Transcription Factor</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Momoeda, M.</creatorcontrib><creatorcontrib>Kawase, M.</creatorcontrib><creatorcontrib>Young, N.S.</creatorcontrib><creatorcontrib>Kajigaya, S.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Protein expression and purification</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Momoeda, M.</au><au>Kawase, M.</au><au>Young, N.S.</au><au>Kajigaya, S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification of Functional Recombinant YY1 Expressed in Sf9 Insect Cells</atitle><jtitle>Protein expression and purification</jtitle><addtitle>Protein Expr Purif</addtitle><date>1995-04-01</date><risdate>1995</risdate><volume>6</volume><issue>2</issue><spage>169</spage><epage>175</epage><pages>169-175</pages><issn>1046-5928</issn><eissn>1096-0279</eissn><abstract>We expressed a gene for YY1, the multifunctional mammalian transcription factor, with a recombinant baculovirus in insect cells and obtained high levels of protein in cell. lysates. A simple and efficient purification method was developed. In the final step, we used DNA affinity chromatography with concatermerized sequence derived from the upstream region of the B19 parvovirus P6 promoter, which has strong affinity for YY1, Approximately 1.8 mg of highly purified YY1 was obtained from 10
9 Sf9 cells. Highly purified YY1 behaved as authentic YY1 on electrophoretic mobility shift assays and in enhancing transcription from the P6 promoter. YY1 produced in a baculovirus system should prove useful for
in vitro transcription assays of YY1 and related regulatory proteins.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>7606165</pmid><doi>10.1006/prep.1995.1021</doi><tpages>7</tpages></addata></record> |
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| ispartof | Protein expression and purification, 1995-04, Vol.6 (2), p.169-175 |
| issn | 1046-5928 1096-0279 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Animals Base Sequence Cell Line Chromatography, Affinity Chromatography, Ion Exchange DNA, Viral - metabolism DNA-Binding Proteins - biosynthesis DNA-Binding Proteins - genetics DNA-Binding Proteins - isolation & purification Erythroid-Specific DNA-Binding Factors Gene Expression Humans Molecular Sequence Data Nucleopolyhedrovirus Parvovirus B19, Human - genetics Promoter Regions, Genetic Recombinant Fusion Proteins - biosynthesis Recombinant Fusion Proteins - isolation & purification Spodoptera Transcription Factors - biosynthesis Transcription Factors - genetics Transcription Factors - isolation & purification YY1 Transcription Factor |
| title | Purification of Functional Recombinant YY1 Expressed in Sf9 Insect Cells |
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