Heterocyst pattern formation controlled by a diffusible peptide

Science

Volumn 282, Issue 5390, 1998, Pages 935-938

  Yoon, Ho Sung ^a   Golden, James W ^a  

^a TEXAS A AND M UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; BACILLUS; BASE PAIRING; CELL COMMUNICATION; CELLULAR DISTRIBUTION; CYANOBACTERIUM; GENE EXPRESSION; MYXOCOCCALES; PHOTOSYNTHESIS; PRIORITY JOURNAL; SIGNAL TRANSDUCTION;

AMINO ACID SEQUENCE; ANABAENA; BACTERIAL PROTEINS; BASE SEQUENCE; COSMIDS; CULTURE MEDIA; DIFFUSION; GENES, BACTERIAL; GENES, REPORTER; GENETIC COMPLEMENTATION TEST; MOLECULAR SEQUENCE DATA; MUTATION, MISSENSE; NITRATES; NITROGEN FIXATION; OLIGOPEPTIDES; PEPTIDE FRAGMENTS; PHENOTYPE; PROMOTER REGIONS (GENETICS); RECOMBINANT FUSION PROTEINS; SIGNAL TRANSDUCTION; TRANSCRIPTION, GENETIC;

EID: 0032582692     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.282.5390.935     Document Type: Article

References (32)

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11. petE-patS fragment from pAM1697 in pAM504 (79). pAM1716 contains patS in the reverse orientation. Two independent clones for each construct were tested, and they produced similar results.
12. + exconjugants were selected for plasmid isolation and DNA sequencing.
13. + exconjugants were selected for plasmid isolation and DNA sequencing.
14. O for 0, 6, 14, 18, 27, and 48 hours and harvested for β-Gal assays. Cells were lysed and assayed as previously described [M. R. Schaefer and S. S. Golden, J. Bacteriol. 171, 3973 (1989)], except that the filaments were frozen at -85°C before processing. β-Gal was measured and expressed as specific activity (nanomoles of o-nitrophenyl-β-D-galactopyranoside per minute per milligram of protein).
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20. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
21. r cassette replaces a 381-bp Eco RV-Sca I fragment containing the entire patS gene. Four independent isolates with identical phenotypes were obtained after selection on media containing sucrose (5%), spectinomycin (2 μg/ml), and streptomycin (2 μg/ml). The structure of the deletion was confirmed by Southern (DNA) blot analysis.
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28. Details of plasmid constructions are available from the authors. Subclones were from pAM1035, which contains patS on a 3.3-kb Bam HI-Cla I fragment that was cloned into the same sites of pBluescript II KS(-). pAM1882, pAM1687, and pAM1835 contain restriction fragments in the conjugal shuttle vector pAM504 (19). pAM1685 contains patS that was amplified by PCR and cloned into pAM504. pAM1686 contains the same insert cloned into pAM505 (identical to pAM504 except that the Bam HI and Sac I sites are reversed). pAM1691 and pAM1695 contain patS cloned into pAM743, which contains the Anabaena PCC 7120 glnA promoter from pAM658 (20) in pAM504. The sequences of all inserts that were generated by PCR were confirmed after subcloning.
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31. rbcL on a shuttle vector, to make pAM1954. pAM1956 contains promoterless gfp from pKEN2-GFPmut2 inserted into pAM505.
32. We thank L. Whorff for contributions to the initial analysis of cosmid 8E11; A. Ott for technical assistance; and W. J. Buikema, J. Elhai, S. S. Golden, M. D. Manson, and members of our laboratory for critically reading the manuscript. Supported in part by NIH grant GM36890.