Cloning of macromomycin apoprotein gene from Streptomyces macromomyceticus by use of 50-mer deoxynucleotide probes.
Identifieur interne : 002049 ( Ncbi/Checkpoint ); précédent : 002048; suivant : 002050Cloning of macromomycin apoprotein gene from Streptomyces macromomyceticus by use of 50-mer deoxynucleotide probes.
Auteurs : M. Hori [Japon] ; N. Sakata ; Y. Niino ; O. Makabe ; M. Hamada ; S. Mizuno ; K. HottaSource :
- The Journal of antibiotics [ 0021-8820 ] ; 1988.
Descripteurs français
- KwdFr :
- Apoprotéines (biosynthèse), Apoprotéines (génétique), Clonage moléculaire, Données de séquences moléculaires, Escherichia coli (génétique), Escherichia coli (métabolisme), Peptides, Plasmides, Protéines bactériennes, Protéines et peptides de signalisation intercellulaire, Régulation de l'expression des gènes, Similitude de séquences d'acides nucléiques, Sondes d'acide nucléique, Streptomyces (génétique), Streptomyces (métabolisme), Séquence d'acides aminés, Séquence nucléotidique, Vecteurs génétiques.
- MESH :
- biosynthèse : Apoprotéines.
- génétique : Apoprotéines, Escherichia coli, Streptomyces.
- métabolisme : Escherichia coli, Streptomyces.
- Clonage moléculaire, Données de séquences moléculaires, Peptides, Plasmides, Protéines bactériennes, Protéines et peptides de signalisation intercellulaire, Régulation de l'expression des gènes, Similitude de séquences d'acides nucléiques, Sondes d'acide nucléique, Séquence d'acides aminés, Séquence nucléotidique, Vecteurs génétiques.
English descriptors
- KwdEn :
- Amino Acid Sequence, Apoproteins (biosynthesis), Apoproteins (genetics), Bacterial Proteins, Base Sequence, Cloning, Molecular, Escherichia coli (genetics), Escherichia coli (metabolism), Gene Expression Regulation, Genetic Vectors, Intercellular Signaling Peptides and Proteins, Molecular Sequence Data, Nucleic Acid Probes, Peptides, Plasmids, Sequence Homology, Nucleic Acid, Streptomyces (genetics), Streptomyces (metabolism).
- MESH :
- chemical , biosynthesis : Apoproteins.
- chemical , genetics : Apoproteins.
- genetics : Escherichia coli, Streptomyces.
- metabolism : Escherichia coli, Streptomyces.
- Amino Acid Sequence, Bacterial Proteins, Base Sequence, Cloning, Molecular, Gene Expression Regulation, Genetic Vectors, Intercellular Signaling Peptides and Proteins, Molecular Sequence Data, Nucleic Acid Probes, Peptides, Plasmids, Sequence Homology, Nucleic Acid.
Abstract
A mixed probe consisting of two synthetic deoxynucleotides (52 and 54 mers referred to as 50-mer) with arbitrarily chosen C or G for the third letters was prepared based on the amino acid sequences No. 31-48 and No. 72-90 of macromomycin (MCM) apoprotein and successfully used to clone the MCM apoprotein gene. Digestion with Sph I of total DNA of MCM-producing Streptomyces macromomyceticus M480-M1 yielded a 2.6-kb fragment that hybridized strongly to the probes. The hybridized probe was stable to washing with 3 x SSC at 75 degrees C. Radioactivity derived from the hybridized probe was comparable to that expected theoretically from hybridization between the probe and the true target sequence. The 2.6-kb fragment was cloned into Escherichia coli RR1 with pBR322 and subsequently subcloned into Streptomyces lividans TK21 with pIJ702. Nucleotide sequence analysis of the cloned fragment verified the existence of the sequence corresponding to the amino acid sequence of MCM apoprotein and about 90% homologies with the probes. Thus, the use of relatively long deoxynucleotide probes with arbitrarily chosen C or G for the third letters will be advantageous in cloning Streptomyces protein genes where more than 90% of the third letters have been known to be C or G. In addition, theoretical diagnosis of hybridization should be a great help to distinguish true positives from false ones.
DOI: 10.7164/antibiotics.41.1462
PubMed: 3056895
Affiliations:
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pubmed:3056895Le document en format XML
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Makabe</name></author><author><name sortKey="Hamada, M" sort="Hamada, M" uniqKey="Hamada M" first="M" last="Hamada">M. Hamada</name></author><author><name sortKey="Mizuno, S" sort="Mizuno, S" uniqKey="Mizuno S" first="S" last="Mizuno">S. Mizuno</name></author><author><name sortKey="Hotta, K" sort="Hotta, K" uniqKey="Hotta K" first="K" last="Hotta">K. 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Hori</name><affiliation wicri:level="3"><nlm:affiliation>Showa College of Pharmaceutical Sciences, Tokyo, Japan.</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>Showa College of Pharmaceutical Sciences, Tokyo</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Sakata, N" sort="Sakata, N" uniqKey="Sakata N" first="N" last="Sakata">N. Sakata</name></author><author><name sortKey="Niino, Y" sort="Niino, Y" uniqKey="Niino Y" first="Y" last="Niino">Y. Niino</name></author><author><name sortKey="Makabe, O" sort="Makabe, O" uniqKey="Makabe O" first="O" last="Makabe">O. Makabe</name></author><author><name sortKey="Hamada, M" sort="Hamada, M" uniqKey="Hamada M" first="M" last="Hamada">M. Hamada</name></author><author><name sortKey="Mizuno, S" sort="Mizuno, S" uniqKey="Mizuno S" first="S" last="Mizuno">S. Mizuno</name></author><author><name sortKey="Hotta, K" sort="Hotta, K" uniqKey="Hotta K" first="K" last="Hotta">K. Hotta</name></author></analytic><series><title level="j">The Journal of antibiotics</title><idno type="ISSN">0021-8820</idno><imprint><date when="1988" type="published">1988</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Apoproteins (biosynthesis)</term><term>Apoproteins (genetics)</term><term>Bacterial Proteins</term><term>Base Sequence</term><term>Cloning, Molecular</term><term>Escherichia coli (genetics)</term><term>Escherichia coli (metabolism)</term><term>Gene Expression Regulation</term><term>Genetic Vectors</term><term>Intercellular Signaling Peptides and Proteins</term><term>Molecular Sequence Data</term><term>Nucleic Acid Probes</term><term>Peptides</term><term>Plasmids</term><term>Sequence Homology, Nucleic Acid</term><term>Streptomyces (genetics)</term><term>Streptomyces (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Apoprotéines (biosynthèse)</term><term>Apoprotéines (génétique)</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Escherichia coli (génétique)</term><term>Escherichia coli (métabolisme)</term><term>Peptides</term><term>Plasmides</term><term>Protéines bactériennes</term><term>Protéines et peptides de signalisation intercellulaire</term><term>Régulation de l'expression des gènes</term><term>Similitude de séquences d'acides nucléiques</term><term>Sondes d'acide nucléique</term><term>Streptomyces (génétique)</term><term>Streptomyces (métabolisme)</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term><term>Vecteurs génétiques</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Apoproteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Apoproteins</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Apoprotéines</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Escherichia coli</term><term>Streptomyces</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Apoprotéines</term><term>Escherichia coli</term><term>Streptomyces</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Escherichia coli</term><term>Streptomyces</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Escherichia coli</term><term>Streptomyces</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Bacterial Proteins</term><term>Base Sequence</term><term>Cloning, Molecular</term><term>Gene Expression Regulation</term><term>Genetic Vectors</term><term>Intercellular Signaling Peptides and Proteins</term><term>Molecular Sequence Data</term><term>Nucleic Acid Probes</term><term>Peptides</term><term>Plasmids</term><term>Sequence Homology, Nucleic Acid</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Peptides</term><term>Plasmides</term><term>Protéines bactériennes</term><term>Protéines et peptides de signalisation intercellulaire</term><term>Régulation de l'expression des gènes</term><term>Similitude de séquences d'acides nucléiques</term><term>Sondes d'acide nucléique</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term><term>Vecteurs génétiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A mixed probe consisting of two synthetic deoxynucleotides (52 and 54 mers referred to as 50-mer) with arbitrarily chosen C or G for the third letters was prepared based on the amino acid sequences No. 31-48 and No. 72-90 of macromomycin (MCM) apoprotein and successfully used to clone the MCM apoprotein gene. Digestion with Sph I of total DNA of MCM-producing Streptomyces macromomyceticus M480-M1 yielded a 2.6-kb fragment that hybridized strongly to the probes. The hybridized probe was stable to washing with 3 x SSC at 75 degrees C. Radioactivity derived from the hybridized probe was comparable to that expected theoretically from hybridization between the probe and the true target sequence. The 2.6-kb fragment was cloned into Escherichia coli RR1 with pBR322 and subsequently subcloned into Streptomyces lividans TK21 with pIJ702. Nucleotide sequence analysis of the cloned fragment verified the existence of the sequence corresponding to the amino acid sequence of MCM apoprotein and about 90% homologies with the probes. Thus, the use of relatively long deoxynucleotide probes with arbitrarily chosen C or G for the third letters will be advantageous in cloning Streptomyces protein genes where more than 90% of the third letters have been known to be C or G. In addition, theoretical diagnosis of hybridization should be a great help to distinguish true positives from false ones.</div></front></TEI><affiliations><list><country><li>Japon</li></country><region><li>Région de Kantō</li></region><settlement><li>Tokyo</li></settlement></list><tree><noCountry><name sortKey="Hamada, M" sort="Hamada, M" uniqKey="Hamada M" first="M" last="Hamada">M. Hamada</name><name sortKey="Hotta, K" sort="Hotta, K" uniqKey="Hotta K" first="K" last="Hotta">K. Hotta</name><name sortKey="Makabe, O" sort="Makabe, O" uniqKey="Makabe O" first="O" last="Makabe">O. Makabe</name><name sortKey="Mizuno, S" sort="Mizuno, S" uniqKey="Mizuno S" first="S" last="Mizuno">S. Mizuno</name><name sortKey="Niino, Y" sort="Niino, Y" uniqKey="Niino Y" first="Y" last="Niino">Y. Niino</name><name sortKey="Sakata, N" sort="Sakata, N" uniqKey="Sakata N" first="N" last="Sakata">N. Sakata</name></noCountry><country name="Japon"><region name="Région de Kantō"><name sortKey="Hori, M" sort="Hori, M" uniqKey="Hori M" first="M" last="Hori">M. Hori</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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