TY - JOUR
T1 - A centromeric DNA sequence colocalized with a centromere-specific histone H3 in tobacco
AU - Nagaki, Kiyotaka
AU - Kashihara, Kazunari
AU - Murata, Minoru
N1 - Funding Information:
Fig. 5 A map of Nt2-7 and Southern blot of genomic DNA. Vertical and horizontal arrows indicate recognition sites of restriction enzymes and primer sites on map (a), respectively. Five micrograms of genomic DNA was digested with AluI (lane 1), HaeIII (lane 2), Sau3AI (lane 3), or KpnI (lane 4). Probes: Nt2-7-1 (b) and Nt2-7-2 (c). d A merged Fig. 6 FISH using the Nt2-7 subclones. Nt2-7-1 (a–c) and Nt2-7-2 (d–f) were hybridized with chromosomes from tobacco root tips. DAPI-stained chromosomes (a, d), FISH signals (b, e), and merged image of chromosomes and FISH signals (c, f). Double-color FISH image of b (red) and c (blue). Overlapped signals were detected as purple signals. e Southern hybridization probed with Nt2-7-2. HaeIII-digested genomic DNA from N. tabaccum (lane 1), N. sylvestris (lane 2), and N. tomentosiformis (lane 3) using Nt2-7-1 (h, red signals in j) and Nt2-7-2 (i, green signals in j). DAPI-stained chromosomes (g), FISH signals (h, i), and merged image of chromosomes and FISH signals (j). Scale bar 10 μm Acknowledgments This work was supported by a grant from the Ministry of Education, Culture, Sports, Science and Technology in Japan (#18770007). Seeds of N. sylvestris and N. tomentosiformis were kindly provided by Japan Tobacco Inc.
Copyright:
Copyright 2009 Elsevier B.V., All rights reserved.
PY - 2009
Y1 - 2009
N2 - Centromeres play an important role in segregating chromosomes into daughter cells, and centromeric DNA assembles specific proteins to form a complex referred to as the kinetochore. Among these proteins, centromere-specific histone H3 (CENH3) is one of the most characterized and found to be located only on active centromeres. We isolated four different CENH3-coding complementary DNAs (cDNAs), two from Nicotiana tabaccum and one each from the ancestral diploid species, Nicotiana sylvestris and Nicotiana tomentosiformis and raised an antibody against N-terminal amino acid sequences deduced from the cDNAs. Immunostaining with the antibody revealed the preferential centromere localization, indicating that the cDNAs cloned in this study encode authentic tobacco CENH3. A tobacco centromeric DNA sequence (Nt2-7) was also identified by chromatin immunoprecipitation cloning using the antibody.
AB - Centromeres play an important role in segregating chromosomes into daughter cells, and centromeric DNA assembles specific proteins to form a complex referred to as the kinetochore. Among these proteins, centromere-specific histone H3 (CENH3) is one of the most characterized and found to be located only on active centromeres. We isolated four different CENH3-coding complementary DNAs (cDNAs), two from Nicotiana tabaccum and one each from the ancestral diploid species, Nicotiana sylvestris and Nicotiana tomentosiformis and raised an antibody against N-terminal amino acid sequences deduced from the cDNAs. Immunostaining with the antibody revealed the preferential centromere localization, indicating that the cDNAs cloned in this study encode authentic tobacco CENH3. A tobacco centromeric DNA sequence (Nt2-7) was also identified by chromatin immunoprecipitation cloning using the antibody.
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U2 - 10.1007/s00412-008-0193-1
DO - 10.1007/s00412-008-0193-1
M3 - Article
C2 - 19048266
AN - SCOPUS:62549159969
SN - 0009-5915
VL - 118
SP - 249
EP - 257
JO - Chromosoma
JF - Chromosoma
IS - 2
ER -