Molecular cloning and characterization of the actin-depolymerizing factor gene in Gossypium barbadense
The Sea Island cotton (Gossypium barbadense L.) has been highly valued in verticillium wilt resistance and many fiber qualities including fiber length, strength and fineness. To identify whether it had some special genes in fiber development in comparison with the Upland cotton (G. hirsutum L.), an...
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| Veröffentlicht in: | Genes & Genetic Systems 2008, Vol.83(5), pp.383-391 |
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| creator | Chi, J.(Agricultural Univ. of Hebei (China)) Wang, X Zhou, H Zhang, G Sun, Y Li, Z Ma, Z |
| description | The Sea Island cotton (Gossypium barbadense L.) has been highly valued in verticillium wilt resistance and many fiber qualities including fiber length, strength and fineness. To identify whether it had some special genes in fiber development in comparison with the Upland cotton (G. hirsutum L.), an actin-depolymerizing factor (ADF) gene was cloned and characterized in this research. A 420 bp open reading frame of the cloned gene, named GbADF1, encoded a protein of 139 amino acids, including 39.57% nonpolar amino acids, 17.27% acidic amino acids, 15.83% basic amino acids and 31.92% hydrophobic amino acids. Its molecular weight was about 15 kDa, and pI 5.04. GbADF1 contained two conserved domains, 6-Ser and the PIP2/actin binding site. Its amino acid sequence was similar to the ADF/cofilin family of other plants. Compared with cDNA sequence, the GbADF1 gene contained one intron near the 3’ end in genomic sequence. Semi-quantitative RT-PCR result showed that GbADF1 was a constitutive expression gene in cotton, and higher expression level was detected in fibers than in trophic tissues. The GbADF1 was successfully expressed as a fusion protein in Escherichia coli BL21 (DE3). The molecular weight was firstly calculated by SDS-PAGE. Western blotting analysis confirmed the existence of a protein corresponding to GbADF1. The structure of GbADF1 was different from that of other ADF genes in higher plant, although the coding sequences of all cloned ADFs were highly conserved. |
| doi_str_mv | 10.1266/ggs.83.383 |
| format | Article |
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To identify whether it had some special genes in fiber development in comparison with the Upland cotton (G. hirsutum L.), an actin-depolymerizing factor (ADF) gene was cloned and characterized in this research. A 420 bp open reading frame of the cloned gene, named GbADF1, encoded a protein of 139 amino acids, including 39.57% nonpolar amino acids, 17.27% acidic amino acids, 15.83% basic amino acids and 31.92% hydrophobic amino acids. Its molecular weight was about 15 kDa, and pI 5.04. GbADF1 contained two conserved domains, 6-Ser and the PIP2/actin binding site. Its amino acid sequence was similar to the ADF/cofilin family of other plants. Compared with cDNA sequence, the GbADF1 gene contained one intron near the 3’ end in genomic sequence. Semi-quantitative RT-PCR result showed that GbADF1 was a constitutive expression gene in cotton, and higher expression level was detected in fibers than in trophic tissues. The GbADF1 was successfully expressed as a fusion protein in Escherichia coli BL21 (DE3). The molecular weight was firstly calculated by SDS-PAGE. Western blotting analysis confirmed the existence of a protein corresponding to GbADF1. The structure of GbADF1 was different from that of other ADF genes in higher plant, although the coding sequences of all cloned ADFs were highly conserved.</description><identifier>ISSN: 1341-7568</identifier><identifier>EISSN: 1880-5779</identifier><identifier>DOI: 10.1266/ggs.83.383</identifier><identifier>PMID: 19168989</identifier><language>eng</language><publisher>Japan: The Genetics Society of Japan</publisher><subject>ACTIN ; Actin Depolymerizing Factors - genetics ; Actin Depolymerizing Factors - metabolism ; ACTINA ; ACTINE ; ADF (actin-depolymerizing factor) ; Base Sequence ; BINDING PROTEINS ; CLONACION MOLECULAR ; CLONAGE MOLECULAIRE ; Cloning, Molecular ; DNA, Complementary - genetics ; DNA, Complementary - metabolism ; Escherichia coli ; FIBRAS ; FIBRE ; FIBRES ; GENE ; GENES ; Genes, Plant ; Genome, Plant ; Gossypium - genetics ; Gossypium - metabolism ; GOSSYPIUM BARBADENSE ; MOLECULAR CLONING ; Molecular Sequence Data ; NUCLEOTIDE SEQUENCE ; Phylogeny ; Plant Proteins - genetics ; Plant Proteins - metabolism ; PROTEINAS AGLUTINANTES ; PROTEINE DE LIAISON ; Reverse Transcriptase Polymerase Chain Reaction ; Sea Island cotton ; SECUENCIA NUCLEOTIDICA ; semi-quantitative RT-PCR ; Sequence Alignment ; SEQUENCE NUCLEOTIDIQUE ; Verticillium ; western blotting</subject><ispartof>Genes & Genetic Systems, 2008, Vol.83(5), pp.383-391</ispartof><rights>2008 by The Genetics Society of Japan</rights><rights>Copyright Japan Science and Technology Agency 2008</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c700t-ceb2bcf36d5ae5a61d03c83adb50e8d3e572c6adbc36b293dc1f1318048726573</citedby><cites>FETCH-LOGICAL-c700t-ceb2bcf36d5ae5a61d03c83adb50e8d3e572c6adbc36b293dc1f1318048726573</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,1877,4010,27902,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19168989$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chi, J.(Agricultural Univ. of Hebei (China))</creatorcontrib><creatorcontrib>Wang, X</creatorcontrib><creatorcontrib>Zhou, H</creatorcontrib><creatorcontrib>Zhang, G</creatorcontrib><creatorcontrib>Sun, Y</creatorcontrib><creatorcontrib>Li, Z</creatorcontrib><creatorcontrib>Ma, Z</creatorcontrib><creatorcontrib>Agricultural University of Hebei</creatorcontrib><creatorcontrib>College of Life Science</creatorcontrib><creatorcontrib>Normal University of Langfang</creatorcontrib><creatorcontrib>Key Laboratory of Crop Germplasm Resources of Hebei Province</creatorcontrib><title>Molecular cloning and characterization of the actin-depolymerizing factor gene in Gossypium barbadense</title><title>Genes & Genetic Systems</title><addtitle>Genes Genet. Syst.</addtitle><description>The Sea Island cotton (Gossypium barbadense L.) has been highly valued in verticillium wilt resistance and many fiber qualities including fiber length, strength and fineness. To identify whether it had some special genes in fiber development in comparison with the Upland cotton (G. hirsutum L.), an actin-depolymerizing factor (ADF) gene was cloned and characterized in this research. A 420 bp open reading frame of the cloned gene, named GbADF1, encoded a protein of 139 amino acids, including 39.57% nonpolar amino acids, 17.27% acidic amino acids, 15.83% basic amino acids and 31.92% hydrophobic amino acids. Its molecular weight was about 15 kDa, and pI 5.04. GbADF1 contained two conserved domains, 6-Ser and the PIP2/actin binding site. Its amino acid sequence was similar to the ADF/cofilin family of other plants. Compared with cDNA sequence, the GbADF1 gene contained one intron near the 3’ end in genomic sequence. Semi-quantitative RT-PCR result showed that GbADF1 was a constitutive expression gene in cotton, and higher expression level was detected in fibers than in trophic tissues. The GbADF1 was successfully expressed as a fusion protein in Escherichia coli BL21 (DE3). The molecular weight was firstly calculated by SDS-PAGE. Western blotting analysis confirmed the existence of a protein corresponding to GbADF1. The structure of GbADF1 was different from that of other ADF genes in higher plant, although the coding sequences of all cloned ADFs were highly conserved.</description><subject>ACTIN</subject><subject>Actin Depolymerizing Factors - genetics</subject><subject>Actin Depolymerizing Factors - metabolism</subject><subject>ACTINA</subject><subject>ACTINE</subject><subject>ADF (actin-depolymerizing factor)</subject><subject>Base Sequence</subject><subject>BINDING PROTEINS</subject><subject>CLONACION MOLECULAR</subject><subject>CLONAGE MOLECULAIRE</subject><subject>Cloning, Molecular</subject><subject>DNA, Complementary - genetics</subject><subject>DNA, Complementary - metabolism</subject><subject>Escherichia coli</subject><subject>FIBRAS</subject><subject>FIBRE</subject><subject>FIBRES</subject><subject>GENE</subject><subject>GENES</subject><subject>Genes, Plant</subject><subject>Genome, Plant</subject><subject>Gossypium - genetics</subject><subject>Gossypium - metabolism</subject><subject>GOSSYPIUM BARBADENSE</subject><subject>MOLECULAR CLONING</subject><subject>Molecular Sequence Data</subject><subject>NUCLEOTIDE SEQUENCE</subject><subject>Phylogeny</subject><subject>Plant Proteins - genetics</subject><subject>Plant Proteins - metabolism</subject><subject>PROTEINAS AGLUTINANTES</subject><subject>PROTEINE DE LIAISON</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Sea Island cotton</subject><subject>SECUENCIA NUCLEOTIDICA</subject><subject>semi-quantitative RT-PCR</subject><subject>Sequence Alignment</subject><subject>SEQUENCE NUCLEOTIDIQUE</subject><subject>Verticillium</subject><subject>western blotting</subject><issn>1341-7568</issn><issn>1880-5779</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNksuL1TAUxosozkM37pWA4GKg1zyaNF2JDM6ojOhC1yFNT3tz6U2uSbu4_vWe0jsOuNFAnueX7yT5UhQvGN0wrtTbYcgbLTZCi0fFOdOalrKum8c4FhUra6n0WXGR845SThstnhZnrGFKN7o5L_ovcQQ3jzYRN8bgw0Bs6Ijb2mTdBMn_spOPgcSeTFsguOZD2cEhjsf9El029LgaExkgAPGB3Macjwc_70lrU2s7CBmeFU96O2Z4fuovix83H75ffyzvvt5-un5_V7qa0ql00PLW9UJ10oK0inVUOC1s10oKuhMga-4UTp1QLW9E51jPBNO00jVXshaXxZtV95DizxnyZPY-OxhHGyDO2SgsnDL-T5CjJGNM_QfIGZNUI_j6L3AX5xTwtoZVqhKV5JwidbVSLuEzJejNIfm9TUfDqFncNOim0cKgmwi_OknO7R66B_RkHwI3K4BR7ywaOPoAD4ndVqDccTkl1QaroBI7YSjKY9MwrXQtKxR6twrt8mQH-JPJpsm7Ee4PJddm2XwfWX6KgYAKL1eF3kZjh-Sz-fwN0zYU01ImfgNyMtEh</recordid><startdate>2008</startdate><enddate>2008</enddate><creator>Chi, J.(Agricultural Univ. of Hebei (China))</creator><creator>Wang, X</creator><creator>Zhou, H</creator><creator>Zhang, G</creator><creator>Sun, Y</creator><creator>Li, Z</creator><creator>Ma, Z</creator><general>The Genetics Society of Japan</general><general>Japan Science and Technology Agency</general><scope>FBQ</scope><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>7SS</scope><scope>7TK</scope><scope>8FD</scope><scope>FR3</scope><scope>K9.</scope><scope>NAPCQ</scope><scope>P64</scope><scope>RC3</scope><scope>7QL</scope><scope>7TM</scope><scope>C1K</scope><scope>M7N</scope><scope>7X8</scope></search><sort><creationdate>2008</creationdate><title>Molecular cloning and characterization of the actin-depolymerizing factor gene in Gossypium barbadense</title><author>Chi, J.(Agricultural Univ. of Hebei (China)) ; Wang, X ; Zhou, H ; Zhang, G ; Sun, Y ; Li, Z ; Ma, Z</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c700t-ceb2bcf36d5ae5a61d03c83adb50e8d3e572c6adbc36b293dc1f1318048726573</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>ACTIN</topic><topic>Actin Depolymerizing Factors - genetics</topic><topic>Actin Depolymerizing Factors - metabolism</topic><topic>ACTINA</topic><topic>ACTINE</topic><topic>ADF (actin-depolymerizing factor)</topic><topic>Base Sequence</topic><topic>BINDING PROTEINS</topic><topic>CLONACION MOLECULAR</topic><topic>CLONAGE MOLECULAIRE</topic><topic>Cloning, Molecular</topic><topic>DNA, Complementary - genetics</topic><topic>DNA, Complementary - metabolism</topic><topic>Escherichia coli</topic><topic>FIBRAS</topic><topic>FIBRE</topic><topic>FIBRES</topic><topic>GENE</topic><topic>GENES</topic><topic>Genes, Plant</topic><topic>Genome, Plant</topic><topic>Gossypium - genetics</topic><topic>Gossypium - metabolism</topic><topic>GOSSYPIUM BARBADENSE</topic><topic>MOLECULAR CLONING</topic><topic>Molecular Sequence Data</topic><topic>NUCLEOTIDE SEQUENCE</topic><topic>Phylogeny</topic><topic>Plant Proteins - genetics</topic><topic>Plant Proteins - metabolism</topic><topic>PROTEINAS AGLUTINANTES</topic><topic>PROTEINE DE LIAISON</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Sea Island cotton</topic><topic>SECUENCIA NUCLEOTIDICA</topic><topic>semi-quantitative RT-PCR</topic><topic>Sequence Alignment</topic><topic>SEQUENCE NUCLEOTIDIQUE</topic><topic>Verticillium</topic><topic>western blotting</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chi, J.(Agricultural Univ. of Hebei (China))</creatorcontrib><creatorcontrib>Wang, X</creatorcontrib><creatorcontrib>Zhou, H</creatorcontrib><creatorcontrib>Zhang, G</creatorcontrib><creatorcontrib>Sun, Y</creatorcontrib><creatorcontrib>Li, Z</creatorcontrib><creatorcontrib>Ma, Z</creatorcontrib><creatorcontrib>Agricultural University of Hebei</creatorcontrib><creatorcontrib>College of Life Science</creatorcontrib><creatorcontrib>Normal University of Langfang</creatorcontrib><creatorcontrib>Key Laboratory of Crop Germplasm Resources of Hebei Province</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Neurosciences Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Premium</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><jtitle>Genes & Genetic Systems</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chi, J.(Agricultural Univ. of Hebei (China))</au><au>Wang, X</au><au>Zhou, H</au><au>Zhang, G</au><au>Sun, Y</au><au>Li, Z</au><au>Ma, Z</au><aucorp>Agricultural University of Hebei</aucorp><aucorp>College of Life Science</aucorp><aucorp>Normal University of Langfang</aucorp><aucorp>Key Laboratory of Crop Germplasm Resources of Hebei Province</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular cloning and characterization of the actin-depolymerizing factor gene in Gossypium barbadense</atitle><jtitle>Genes & Genetic Systems</jtitle><addtitle>Genes Genet. Syst.</addtitle><date>2008</date><risdate>2008</risdate><volume>83</volume><issue>5</issue><spage>383</spage><epage>391</epage><pages>383-391</pages><issn>1341-7568</issn><eissn>1880-5779</eissn><abstract>The Sea Island cotton (Gossypium barbadense L.) has been highly valued in verticillium wilt resistance and many fiber qualities including fiber length, strength and fineness. To identify whether it had some special genes in fiber development in comparison with the Upland cotton (G. hirsutum L.), an actin-depolymerizing factor (ADF) gene was cloned and characterized in this research. A 420 bp open reading frame of the cloned gene, named GbADF1, encoded a protein of 139 amino acids, including 39.57% nonpolar amino acids, 17.27% acidic amino acids, 15.83% basic amino acids and 31.92% hydrophobic amino acids. Its molecular weight was about 15 kDa, and pI 5.04. GbADF1 contained two conserved domains, 6-Ser and the PIP2/actin binding site. Its amino acid sequence was similar to the ADF/cofilin family of other plants. Compared with cDNA sequence, the GbADF1 gene contained one intron near the 3’ end in genomic sequence. Semi-quantitative RT-PCR result showed that GbADF1 was a constitutive expression gene in cotton, and higher expression level was detected in fibers than in trophic tissues. The GbADF1 was successfully expressed as a fusion protein in Escherichia coli BL21 (DE3). The molecular weight was firstly calculated by SDS-PAGE. Western blotting analysis confirmed the existence of a protein corresponding to GbADF1. The structure of GbADF1 was different from that of other ADF genes in higher plant, although the coding sequences of all cloned ADFs were highly conserved.</abstract><cop>Japan</cop><pub>The Genetics Society of Japan</pub><pmid>19168989</pmid><doi>10.1266/ggs.83.383</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | ACTIN Actin Depolymerizing Factors - genetics Actin Depolymerizing Factors - metabolism ACTINA ACTINE ADF (actin-depolymerizing factor) Base Sequence BINDING PROTEINS CLONACION MOLECULAR CLONAGE MOLECULAIRE Cloning, Molecular DNA, Complementary - genetics DNA, Complementary - metabolism Escherichia coli FIBRAS FIBRE FIBRES GENE GENES Genes, Plant Genome, Plant Gossypium - genetics Gossypium - metabolism GOSSYPIUM BARBADENSE MOLECULAR CLONING Molecular Sequence Data NUCLEOTIDE SEQUENCE Phylogeny Plant Proteins - genetics Plant Proteins - metabolism PROTEINAS AGLUTINANTES PROTEINE DE LIAISON Reverse Transcriptase Polymerase Chain Reaction Sea Island cotton SECUENCIA NUCLEOTIDICA semi-quantitative RT-PCR Sequence Alignment SEQUENCE NUCLEOTIDIQUE Verticillium western blotting |
| title | Molecular cloning and characterization of the actin-depolymerizing factor gene in Gossypium barbadense |
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