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Cloning, Purification, and Polymerization of Capsicum annuum Recombinant α and β Tubulin
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Jang, Myung-Hyun | - |
dc.contributor.author | Kim, Jungmok | - |
dc.contributor.author | Kalme, Satish | - |
dc.contributor.author | Han, Jin-Wook | - |
dc.contributor.author | Yoo, Han Sang | - |
dc.contributor.author | Kim, Jinheung | - |
dc.contributor.author | Koo, Bon-sung | - |
dc.contributor.author | Kim, Sung-Kun | - |
dc.contributor.author | Yoon, Moon-Young | - |
dc.date.accessioned | 2009-08-26T22:31:29Z | - |
dc.date.available | 2009-08-26T22:31:29Z | - |
dc.date.issued | 2008-04-07 | - |
dc.identifier.citation | Biosci. Biotechnol. Biochem., 72, 1048-1055 (2008) | en |
dc.identifier.issn | 0916-8451 (print) | - |
dc.identifier.issn | 1347-6947 (online) | - |
dc.identifier.uri | https://hdl.handle.net/10371/7872 | - |
dc.description.abstract | α and β tubulin genes were cloned from the Capsicum annuum leaves using rapid amplification of cDNA ends (RACE)-PCR. Nucleotide sequence analysis revealed that 1,353 bp Capsicum annuum α⁄β-tubulin (CAnm α⁄β-TUB) encodes a protein of 450 amino acids (aa) each. The recombinant α⁄β tubulin was overexpressed mainly as an inclusion body in Escherichia coli BL21 (DE3), upon induction with 0.2 mM isopropyl-β-D-thiogalactopyranoside (IPTG), and its content was as high as 50% of the total protein content. Effective fusion protein purification and refolding are described. The average yields of α and β tubulin were 2.0 and 1.3 mg/l of culture respectively. The apparent molecular weight of each tubulin was estimated to be 55 kDa by SDS-polyacrylamide gel electrophoresis (PAGE). The tubulin monomers were found to be assembly competent using a standard dimerization assay, and also retained antigenicity with anti-His/T7 antibodies. The purified tubulins were polymerized to microtubule-like structures in the presence of 2 mM guanosine 5′-triphosphate (GTP). | en |
dc.description.sponsorship | This study was carried out with the support of the
On-Site Cooperative Agriculture Research Project (20070201080024) and the National Institute of Agricultural Biotechnology, RDA, of the Republic of Korea. | en |
dc.language.iso | en | en |
dc.publisher | Japan Society for Bioscience, Biotechnology, and Agrochemistry | en |
dc.subject | Capsicum annuum | en |
dc.subject | dimerization | en |
dc.subject | microtubule | en |
dc.subject | pepper | en |
dc.title | Cloning, Purification, and Polymerization of Capsicum annuum Recombinant α and β Tubulin | en |
dc.type | Article | en |
dc.contributor.AlternativeAuthor | 장명현 | - |
dc.contributor.AlternativeAuthor | 김정목 | - |
dc.contributor.AlternativeAuthor | 한진욱 | - |
dc.contributor.AlternativeAuthor | 유한상 | - |
dc.contributor.AlternativeAuthor | 김진흥 | - |
dc.contributor.AlternativeAuthor | 구본성 | - |
dc.contributor.AlternativeAuthor | 김성근 | - |
dc.contributor.AlternativeAuthor | 윤무영 | - |
dc.identifier.doi | 10.1271/bbb.70794 | - |
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