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Cloning, Purification, and Polymerization of Capsicum annuum Recombinant α and β Tubulin

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dc.contributor.authorJang, Myung-Hyun-
dc.contributor.authorKim, Jungmok-
dc.contributor.authorKalme, Satish-
dc.contributor.authorHan, Jin-Wook-
dc.contributor.authorYoo, Han Sang-
dc.contributor.authorKim, Jinheung-
dc.contributor.authorKoo, Bon-sung-
dc.contributor.authorKim, Sung-Kun-
dc.contributor.authorYoon, Moon-Young-
dc.date.accessioned2009-08-26T22:31:29Z-
dc.date.available2009-08-26T22:31:29Z-
dc.date.issued2008-04-07-
dc.identifier.citationBiosci. Biotechnol. Biochem., 72, 1048-1055 (2008)en
dc.identifier.issn0916-8451 (print)-
dc.identifier.issn1347-6947 (online)-
dc.identifier.urihttps://hdl.handle.net/10371/7872-
dc.description.abstractα 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.sponsorshipThis 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.isoenen
dc.publisherJapan Society for Bioscience, Biotechnology, and Agrochemistryen
dc.subjectCapsicum annuumen
dc.subjectdimerizationen
dc.subjectmicrotubuleen
dc.subjectpepperen
dc.titleCloning, Purification, and Polymerization of Capsicum annuum Recombinant α and β Tubulinen
dc.typeArticleen
dc.contributor.AlternativeAuthor장명현-
dc.contributor.AlternativeAuthor김정목-
dc.contributor.AlternativeAuthor한진욱-
dc.contributor.AlternativeAuthor유한상-
dc.contributor.AlternativeAuthor김진흥-
dc.contributor.AlternativeAuthor구본성-
dc.contributor.AlternativeAuthor김성근-
dc.contributor.AlternativeAuthor윤무영-
dc.identifier.doi10.1271/bbb.70794-
Appears in Collections:
College of Veterinary Medicine (수의과대학)Dept. of Veterinary Medicine (수의학과)Journal Papers (저널논문_수의학과)
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