Tobacco mitotic cyclins: cloning, characterization, gene expression and functional assay.
Three cyclin cDNA clones (Ntcyc25, Ntcyc27, Ntcyc29) have been isolated from tobacco (Nicotiana tabacum) using the PCR cloning method. The encoded Ntcyc cyclins were highly homologous to mitotic cyclins. In synchronized tobacco suspension cultured cells, the mRNA levels of Ntcyc25 and Ntcyc27 were detectable through S, G2 and M phases, while the Ntcyc29 mRNA was detected from G2 to M phase. These expression patterns classified the Ntcyc25 and Ntcyc27 into A-type cyclin and Ntcyc29 into B-type cyclin. The three genes were expressed in growing tobacco cultured cells but ceased to be expressed when cells entered the stationary phase, indicating that the expression of these cyclin genes was well correlated with cell growth. The N-terminal truncated Ntcyc25 and Ntcyc29 cDNAs were able to rescue G1 cyclin mutant of Saccharomyces cerevisiae, while the full-length of Ntcyc27 protein could also partially perform G1 cyclin functions.
Setiady
Y Y
YY
Department of Biotechnology, Faculty of Engineering, Osaka University, Japan.
Sekine
M
M
Hariguchi
N
N
Yamamoto
T
T
Kouchi
H
H
Shinmyo
A
A
D000595:Amino Acid Sequence; D016590:Aphidicolin / Q000494:pharmacology; D001483:Base Sequence; D015152:Blotting, Northern; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology; D003001:Cloning, Molecular; D016213:Cyclins / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary / Q000302:isolation & purification; D015870:Gene Expression; D017343:Genes, Plant; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D016133:Polymerase Chain Reaction; D018749:RNA, Plant / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014026:Nicotiana / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*
D016213:Cyclins; D017931:DNA Primers; D018076:DNA, Complementary; D018749:RNA, Plant; D011994:Recombinant Proteins; D016590:Aphidicolin