PMID- 19394244
OWN - NLM
STAT- MEDLINE
DCOM- 20090910
LR  - 20200930
IS  - 1873-2690 (Electronic)
IS  - 0981-9428 (Linking)
VI  - 47
IP  - 9
DP  - 2009 Sep
TI  - Molecular cloning and biochemical characterization of alpha- and beta-tubulin 
      from potato plants (Solanum tuberosum L.).
PG  - 761-8
LID - 10.1016/j.plaphy.2009.04.001 [doi]
AB  - Few studies have investigated microtubules from plants that host pathogenic 
      fungi. Considerable efforts are underway to find an antimitotic agent against 
      plant pathogens like Phytophthora infestans. However, screening the effects of 
      antifungal agents on plant tubulin in vivo or using purified native microtubule 
      in vitro is a time consuming process. A recombinant, correctly folded, 
      microtubule-like structure forming tubulin could accelerate research in this 
      area. In this study, we cloned full length cDNAs isolated from potato leaves 
      using reverse-transcribed polymerase chain reaction (RT-PCR). Solanum tuberosum 
      (Stub) alpha-tubulin and beta-tubulin were predicted to encode 449 and 451 amino 
      acid long proteins with molecular masses of 57 kDa and 60 kDa, respectively. 
      Average yields of alpha- and beta-tubulin were 2.0-3.5 mg l(-1) and 1.3-3.0 mg 
      l(-1) of culture, respectively. The amino acids, His6, Glu198, and Phe170 
      involved in benomyl sensitivity were conserved in Stub tubulin. The dimerization 
      of tubulin monomers was confirmed by western blot analysis. When combined under 
      appropriate conditions, these recombinant alpha- and beta-tubulins were capable 
      of polymerizing into microtubules. Accessibility of cysteine residues of tubulin 
      revealed that important ligand binding sites were folded correctly. This 
      recombinant tubulin could serve as a control of phytotoxicity of selected 
      antimitotic fungicide compounds during in vitro screening experiments.
FAU - Koo, Bon-Sung
AU  - Koo BS
AD  - Fermentation and Food Processing Division, Department of Korean Food Research for 
      Globalization, National Academy of Agricultural Science, Suwon 441 857, South 
      Korea.
FAU - Kalme, Satish
AU  - Kalme S
FAU - Yeo, Soo-Hwan
AU  - Yeo SH
FAU - Lee, Su-Jae
AU  - Lee SJ
FAU - Yoon, Moon-Young
AU  - Yoon MY
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090409
PL  - France
TA  - Plant Physiol Biochem
JT  - Plant physiology and biochemistry : PPB
JID - 9882449
RN  - 0 (Tubulin)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Cloning, Molecular
MH  - Computational Biology
MH  - Cysteine/genetics
MH  - Dimerization
MH  - Genes, Plant
MH  - Microtubules/metabolism
MH  - Mitosis
MH  - Molecular Sequence Data
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sequence Homology, Amino Acid
MH  - Software
MH  - Solanum tuberosum/*genetics
MH  - Tubulin/*genetics
EDAT- 2009/04/28 09:00
MHDA- 2009/09/11 06:00
CRDT- 2009/04/28 09:00
PHST- 2008/09/30 00:00 [received]
PHST- 2009/03/30 00:00 [revised]
PHST- 2009/04/01 00:00 [accepted]
PHST- 2009/04/28 09:00 [entrez]
PHST- 2009/04/28 09:00 [pubmed]
PHST- 2009/09/11 06:00 [medline]
AID - S0981-9428(09)00104-1 [pii]
AID - 10.1016/j.plaphy.2009.04.001 [doi]
PST - ppublish
SO  - Plant Physiol Biochem. 2009 Sep;47(9):761-8. doi: 10.1016/j.plaphy.2009.04.001. 
      Epub 2009 Apr 9.