PMID- 18391467
OWN - NLM
STAT- MEDLINE
DCOM- 20080801
LR  - 20080424
IS  - 1347-6947 (Electronic)
IS  - 0916-8451 (Linking)
VI  - 72
IP  - 4
DP  - 2008 Apr
TI  - Cloning, purification, and polymerization of Capsicum annuum recombinant alpha 
      and beta tubulin.
PG  - 1048-55
AB  - Alpha and beta 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 alpha/beta-tubulin (CAnm alpha/beta-TUB) 
      encodes a protein of 450 amino acids (aa) each. The recombinant alpha/beta 
      tubulin was overexpressed mainly as an inclusion body in Escherichia coli BL21 
      (DE3), upon induction with 0.2 mM isopropyl-beta-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 alpha and 
      beta 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).
FAU - Jang, Myung-Hyun
AU  - Jang MH
AD  - Department of Chemistry, Hanyang University, Seoul 133-791, Korea.
FAU - Kim, Jungmok
AU  - Kim J
FAU - Kalme, Satish
AU  - Kalme S
FAU - Han, Jin-Wook
AU  - Han JW
FAU - Yoo, Han-Sang
AU  - Yoo HS
FAU - Kim, Jinheung
AU  - Kim J
FAU - Koo, Bon-sung
AU  - Koo BS
FAU - Kim, Sung-Kun
AU  - Kim SK
FAU - Yoon, Moon-Young
AU  - Yoon MY
LA  - eng
SI  - GENBANK/EF495257
SI  - GENBANK/EF495259
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20080407
PL  - England
TA  - Biosci Biotechnol Biochem
JT  - Bioscience, biotechnology, and biochemistry
JID - 9205717
RN  - 0 (Biopolymers)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tubulin)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Biopolymers/chemistry/genetics/isolation & purification/metabolism
MH  - Capsicum/*genetics/*metabolism
MH  - Cloning, Molecular
MH  - Conserved Sequence
MH  - Dimerization
MH  - Gene Expression Regulation, Plant
MH  - Molecular Sequence Data
MH  - Protein Binding
MH  - Protein Structure, Quaternary
MH  - Recombinant Proteins/chemistry/*genetics/isolation & purification/*metabolism
MH  - Sequence Alignment
MH  - Sequence Analysis, Protein
MH  - Sequence Homology, Amino Acid
MH  - Tubulin/chemistry/*genetics/isolation & purification/*metabolism
EDAT- 2008/04/09 09:00
MHDA- 2008/08/02 09:00
CRDT- 2008/04/09 09:00
PHST- 2008/04/09 09:00 [pubmed]
PHST- 2008/08/02 09:00 [medline]
PHST- 2008/04/09 09:00 [entrez]
AID - JST.JSTAGE/bbb/70794 [pii]
AID - 10.1271/bbb.70794 [doi]
PST - ppublish
SO  - Biosci Biotechnol Biochem. 2008 Apr;72(4):1048-55. doi: 10.1271/bbb.70794. Epub 
      2008 Apr 7.