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Volumn 277, Issue 5325, 1997, Pages 574-578

Mitosis in living budding yeast: Anaphase a but no metaphase plate

Author keywords

[No Author keywords available]

Indexed keywords

ALPHA TUBULIN; GREEN FLUORESCENT PROTEIN; HYBRID PROTEIN;

EID: 0030847538     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5325.574     Document Type: Article
Times cited : (326)

References (27)
  • 7
    • 1842286844 scopus 로고    scopus 로고
    • note
    • The GFP-Tub1 fusion under the control of the HIS3 promoter was constructed by ligating the entire genomic TUB1 locus with Bam HI and Not I linkers into pAFS78 (8). The resultant GFP-Tub1 fusion was cloned into the Kpn I to Xho I sites of pRS306 to yield pAFS91 and integrated at the URA3 locus after Stu I digestion. The GFP-tubulin fusion was functional because it rescues a normally lethal TUB1 deletion. Strain AFS403 was constructed by transforming strain AFS173 (8) with Stu I-digested pAFS91.
  • 10
    • 1842364155 scopus 로고    scopus 로고
    • note
    • 1 by treatment with α-factor (10 μg/ml) for 3 hours, released from this arrest, induced to express the GFP-Tub1 and GFP-LacI fusions for 30 min (8), and mounted in histidine-free medium. Every 26 s, 12 optical sections (100-ms exposure time) were collected (vertical separation, 0.5 μm) by wide-field deconvolution 3D microscopy (11) with a 60× 1.4 numerical aperture lens.
  • 26
    • 1842286724 scopus 로고    scopus 로고
    • note
    • Strain AFS412 was constructed by integrating pAFS52 (8) at TRP1 near the centromere of chromosome IV and then integrating pAFS114 near the telomere of chromosome IV. Plasmid pAFS114 contains a Kpn I-Sac I fragment of chromosome IV spanning nucleotides 1,436,502 to 1,438,325 in pAFS59 (8) and was integrated into chromosome IV after digestion with Bsa BI.
  • 27
    • 1842311290 scopus 로고    scopus 로고
    • note
    • We thank members of the Murray laboratory; D. O. Morgan and A. Farrell for critical review of the manuscript; A. Desai, T. J. Mitchison, and H. Funabiki for insightful comments and discussion; and J. Fung and S. Parmelee for assistance during data analysis. Supported by grants from NIH and the Packard Foundation.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.