An M13 vector library for cloning DNA with four nucleotide 3′ overhangs
A flexible M13 vector library incorporating the BstXI site has been developed. DNA cut by any currently commercially available restriction endonuclease that generates a 4-nucleotide (nt) 3′ overhang can be ligated into a specific clone of the library. The BstXI enzyme recognizes a 6-bp bipartite pal...
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| Veröffentlicht in: | Plasmid 1990-07, Vol.24 (1), p.68-73 |
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| creator | Panchmatia, F. Kay Cramb, Elizabeth M. Li, Vincent Allen, Frances L. Glickman, Barry W. Waye, Mary M.Y. |
| description | A flexible M13 vector library incorporating the
BstXI site has been developed. DNA cut by any currently commercially available restriction endonuclease that generates a 4-nucleotide (nt) 3′ overhang can be ligated into a specific clone of the library. The
BstXI enzyme recognizes a 6-bp bipartite palindromic sequence. The central nucleotides are not specified, and form a 4-base, 3′ overhang when cut by
BstXI. 5′ CCANNNNN NTGG GGTN NNNNNACC 5′ Since the 4-base overhang formed is not part of the
BstXI recognition sequence, it is possible to generate a library of 256 different clones by introducing the
BstXI site. 151 of the possible 256-member library have been isolated, including all 13 M13BF clones in which the overhang formed by
BstXI digestion is complementary to those formed by currently available restriction endonucleases. Of these 13 vectors,
BstXI digestion of six clones results in nonpalindromic cohesive ends and should facilitate
in vitro tandem gene amplification. The
BstXI site is adjacent to the four codons corresponding to the factor Xa recognition sequence. Hence the vector library could facilitate the expression of a fusion protein that could be proteolytically cleaved by factor Xa. |
| doi_str_mv | 10.1016/0147-619X(90)90026-9 |
| format | Article |
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BstXI site has been developed. DNA cut by any currently commercially available restriction endonuclease that generates a 4-nucleotide (nt) 3′ overhang can be ligated into a specific clone of the library. The
BstXI enzyme recognizes a 6-bp bipartite palindromic sequence. The central nucleotides are not specified, and form a 4-base, 3′ overhang when cut by
BstXI. 5′ CCANNNNN NTGG GGTN NNNNNACC 5′ Since the 4-base overhang formed is not part of the
BstXI recognition sequence, it is possible to generate a library of 256 different clones by introducing the
BstXI site. 151 of the possible 256-member library have been isolated, including all 13 M13BF clones in which the overhang formed by
BstXI digestion is complementary to those formed by currently available restriction endonucleases. Of these 13 vectors,
BstXI digestion of six clones results in nonpalindromic cohesive ends and should facilitate
in vitro tandem gene amplification. The
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BstXI site has been developed. DNA cut by any currently commercially available restriction endonuclease that generates a 4-nucleotide (nt) 3′ overhang can be ligated into a specific clone of the library. The
BstXI enzyme recognizes a 6-bp bipartite palindromic sequence. The central nucleotides are not specified, and form a 4-base, 3′ overhang when cut by
BstXI. 5′ CCANNNNN NTGG GGTN NNNNNACC 5′ Since the 4-base overhang formed is not part of the
BstXI recognition sequence, it is possible to generate a library of 256 different clones by introducing the
BstXI site. 151 of the possible 256-member library have been isolated, including all 13 M13BF clones in which the overhang formed by
BstXI digestion is complementary to those formed by currently available restriction endonucleases. Of these 13 vectors,
BstXI digestion of six clones results in nonpalindromic cohesive ends and should facilitate
in vitro tandem gene amplification. The
BstXI site is adjacent to the four codons corresponding to the factor Xa recognition sequence. Hence the vector library could facilitate the expression of a fusion protein that could be proteolytically cleaved by factor Xa.</description><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cloning, Molecular</subject><subject>Deoxyribonucleases, Type II Site-Specific</subject><subject>DNA, Bacterial - genetics</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Library</subject><subject>Genes, Bacterial</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Genetic Vectors</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular Sequence Data</subject><subject>phage M13</subject><subject>Vectors (cloning, transfer, expression). 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Technologies</topic><topic>Molecular Sequence Data</topic><topic>phage M13</topic><topic>Vectors (cloning, transfer, expression). Insertion sequences and transposons</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Panchmatia, F. Kay</creatorcontrib><creatorcontrib>Cramb, Elizabeth M.</creatorcontrib><creatorcontrib>Li, Vincent</creatorcontrib><creatorcontrib>Allen, Frances L.</creatorcontrib><creatorcontrib>Glickman, Barry W.</creatorcontrib><creatorcontrib>Waye, Mary M.Y.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Plasmid</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Panchmatia, F. 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BstXI site has been developed. DNA cut by any currently commercially available restriction endonuclease that generates a 4-nucleotide (nt) 3′ overhang can be ligated into a specific clone of the library. The
BstXI enzyme recognizes a 6-bp bipartite palindromic sequence. The central nucleotides are not specified, and form a 4-base, 3′ overhang when cut by
BstXI. 5′ CCANNNNN NTGG GGTN NNNNNACC 5′ Since the 4-base overhang formed is not part of the
BstXI recognition sequence, it is possible to generate a library of 256 different clones by introducing the
BstXI site. 151 of the possible 256-member library have been isolated, including all 13 M13BF clones in which the overhang formed by
BstXI digestion is complementary to those formed by currently available restriction endonucleases. Of these 13 vectors,
BstXI digestion of six clones results in nonpalindromic cohesive ends and should facilitate
in vitro tandem gene amplification. The
BstXI site is adjacent to the four codons corresponding to the factor Xa recognition sequence. Hence the vector library could facilitate the expression of a fusion protein that could be proteolytically cleaved by factor Xa.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>2270230</pmid><doi>10.1016/0147-619X(90)90026-9</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
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| language | eng |
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| source | Elsevier ScienceDirect Journals Complete - AutoHoldings; MEDLINE |
| subjects | Base Sequence Biological and medical sciences Biotechnology Cloning, Molecular Deoxyribonucleases, Type II Site-Specific DNA, Bacterial - genetics Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Gene Library Genes, Bacterial Genetic engineering Genetic technics Genetic Vectors Methods. Procedures. Technologies Molecular Sequence Data phage M13 Vectors (cloning, transfer, expression). Insertion sequences and transposons |
| title | An M13 vector library for cloning DNA with four nucleotide 3′ overhangs |
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