NING De-Gang, XU Xu-Dong. A NOVEL REGION INVOLVED IN HETEROCYST DEVELOPMENT AND PATTERN FORMATION IN THE GENOME OFANABAENA SP. PCC 7120[J]. ACTA HYDROBIOLOGICA SINICA, 2004, 28(1): 1-6.
Citation: NING De-Gang, XU Xu-Dong. A NOVEL REGION INVOLVED IN HETEROCYST DEVELOPMENT AND PATTERN FORMATION IN THE GENOME OFANABAENA SP. PCC 7120[J]. ACTA HYDROBIOLOGICA SINICA, 2004, 28(1): 1-6.

A NOVEL REGION INVOLVED IN HETEROCYST DEVELOPMENT AND PATTERN FORMATION IN THE GENOME OFANABAENA SP. PCC 7120

  • Received Date: April 01, 2003
  • Rev Recd Date: June 30, 2003
  • Published Date: January 24, 2004
  • Anabaena sp. PCC 7120 was mutagenized with trans poson Tn5-1087b and screened for mutants unable to grow on dinitrogen and alter ed in heterocyst pattern. Mutant #1801 forms no visible heterocyst within 24h after nitrogen stepdown, and a small number of mature heterocysts within 48h, which a mount to 2.8% of total cell number. The mutant is significantly different from the wild type in rate and frequency of heterocyst differentiation. The flanking g enomic DNA and the transposon were retrieved from the mutant by restriction of i ts total DNA with ClaI, recircularization and electroporation intoE.coli.A s shown with sequencing, the transposon was inserted between Nt. 148 and 149 of alr0099, a gene of unknown function. To prove that the phenotype of the muta nt was indeed due to the insertion withinalr0099 but not a secondary mutati on, we inserted C. K2 cassette, a neomycin resistance marker, into the cyanobact erial genome at the EcoRV site ofalr0099 via double h omologous recombinatio ns. The reconstructed mutant DR214 showed the same phenotype as #1801. These res ults indicate thatalr0099 or its downstream genes sta nd for a novel region involved in heterocyst development and pattern formation in the genome ofAnab aena sp. PCC 7120.
  • [1]
    Wolk C P. Heterocyst formation in Anabaena [C]. Brun Y V, Shimkets L J. Prokaryotic Development. Washington DC:American Society for Microbiology, 2000. 83-104[2] Kong R Q, Xu XD, Wang YQ. Molecular genetics of cyanobacteria:the decade after last review [J]. Acta Hybiologica Sinica, 2001. 25:620-630[孔任秋,徐旭东,王业勤. 蓝藻分子遗传学又十年. 水生生物学报,2001. 25:620-630][3] Buikema W J, Haselkorn R. Expression of the Anabaena hetR gene from a copper-regulated promoter leads to heterocyst differentiation under repressing conditions [J]. Proc Natl Acad Sci USA,2001. 98:2729-2734[4] Yoon H Y, Golden J W. Heterocyst pattern formation controlled by a diffusible peptide [J]. Science, 1998.282:935-938[5] Mcallahan S, Buikema W J. The role of HetN in maintainance of the hetrocyst pattern of Anabaena sp. PCC 7120 [J]. Molecular Microbiology, 2001. 40:941-950[6] Liang J. Scappino L, Haselkorn R. The patA gene product, which contains a region similar to CheY of Escherichia coli, controls heterocyst pattern formation in the cyanobacterium Anabaena 7120 [J]. Proc Natl Acad Sci USA,1992. 89:5655-5659[7] Liang J, Scappino L, and Haselkorn R. The patB gene product, required for growth of the cyanobacterium Anabaena sp. PCC 7120 under nitrogen-limiting conditions, contains ferredoxin and helix-turn-helix domains [J]. J Bacteriol, 1993. 175:1697-1704[8] Elhai J, Wolk C P. Conjugal transfer of DNA to Cyanobacteria [J].Methods. Enzymol, 1988. 167:747-754[9] Ernst A, Black T, Cai Y, et al. Synthesis of nitrogenase in mutants of the cyanobacterium Anabaena sp. strain PCC 7120 affected in heterocyst development or metabolism [J]. J Bacteriol, 1992. 174:6025-6032[10] Hebbar P B, Curtis S E.Characterization of devH, a gene encoding a putative DNA binding protein required for heterocyst function in Anabaena sp. strain PCC 7120[J]. J Bacteriol, 2000. 182:3572-3581[11] Xu XD, Wang YQ, Li SH. Construction of biphasic CAT promoter probe vector shuttling between Anabaena (blue-green lagae) and Escherichia coli [J].Journal of Graduate School, Academia Sinica,1993,10:203-209[徐旭东,王业勤,黎尚豪. 鱼腥藻--大肠杆菌启动子CAT探测载体的构建. 中国科学院研究生院学报, 1993, 10:203-209][12] Elhai J, Wolk C P. A versatile class of positive-selection vectors based on the nonviability of palindrome-containing plasmids that allows the cloning into long polylinkers [J]. Gene, 68:119-138[13] Cai Y, Wolk C P. Use of a conditionally lethal gene in Anabaena sp.strain PCC 7120 to select for double recombinants and to entrap insertion sequences [J]. J Bacteriol, 1990. 172:3138-3145[14] Wolk C P. Physiological basis of the pattern of vegetative growth of ablue-green alga[J].Proc Natl Acad Sci USA,1967,57:1246-1251[15] Wilcox M, Mitchison G J, Smith R J. Pattern formation in the blue-green alga, Anabeana. I. Basic mechanisms [J]. J Cell Sci,1973.12:707-725[16] Wolk C P, Quine M P. Formation of one-dimensional patterns by stochastic processes and by filamentous blue-green algae [J].Develop Biol,1975. 46:370-382[17] Black T A, Cai Y, Wolk C P. Spatial expression and autoregulation of hetR, a gene involved in the control of heterocyst development in Anabaena[J].Mol Microbiol, 1993. 9:77-84

    Wolk C P. Heterocyst formation in Anabaena [C]. Brun Y V, Shimkets L J. Prokaryotic Development. Washington DC:American Society for Microbiology, 2000. 83-104[2] Kong R Q, Xu XD, Wang YQ. Molecular genetics of cyanobacteria:the decade after last review [J]. Acta Hybiologica Sinica, 2001. 25:620-630[孔任秋,徐旭东,王业勤. 蓝藻分子遗传学又十年. 水生生物学报,2001. 25:620-630][3] Buikema W J, Haselkorn R. Expression of the Anabaena hetR gene from a copper-regulated promoter leads to heterocyst differentiation under repressing conditions [J]. Proc Natl Acad Sci USA,2001. 98:2729-2734[4] Yoon H Y, Golden J W. Heterocyst pattern formation controlled by a diffusible peptide [J]. Science, 1998.282:935-938[5] Mcallahan S, Buikema W J. The role of HetN in maintainance of the hetrocyst pattern of Anabaena sp. PCC 7120 [J]. Molecular Microbiology, 2001. 40:941-950[6] Liang J. Scappino L, Haselkorn R. The patA gene product, which contains a region similar to CheY of Escherichia coli, controls heterocyst pattern formation in the cyanobacterium Anabaena 7120 [J]. Proc Natl Acad Sci USA,1992. 89:5655-5659[7] Liang J, Scappino L, and Haselkorn R. The patB gene product, required for growth of the cyanobacterium Anabaena sp. PCC 7120 under nitrogen-limiting conditions, contains ferredoxin and helix-turn-helix domains [J]. J Bacteriol, 1993. 175:1697-1704[8] Elhai J, Wolk C P. Conjugal transfer of DNA to Cyanobacteria [J].Methods. Enzymol, 1988. 167:747-754[9] Ernst A, Black T, Cai Y, et al. Synthesis of nitrogenase in mutants of the cyanobacterium Anabaena sp. strain PCC 7120 affected in heterocyst development or metabolism [J]. J Bacteriol, 1992. 174:6025-6032[10] Hebbar P B, Curtis S E.Characterization of devH, a gene encoding a putative DNA binding protein required for heterocyst function in Anabaena sp. strain PCC 7120[J]. J Bacteriol, 2000. 182:3572-3581[11] Xu XD, Wang YQ, Li SH. Construction of biphasic CAT promoter probe vector shuttling between Anabaena (blue-green lagae) and Escherichia coli [J].Journal of Graduate School, Academia Sinica,1993,10:203-209[徐旭东,王业勤,黎尚豪. 鱼腥藻--大肠杆菌启动子CAT探测载体的构建. 中国科学院研究生院学报, 1993, 10:203-209][12] Elhai J, Wolk C P. A versatile class of positive-selection vectors based on the nonviability of palindrome-containing plasmids that allows the cloning into long polylinkers [J]. Gene, 68:119-138[13] Cai Y, Wolk C P. Use of a conditionally lethal gene in Anabaena sp.strain PCC 7120 to select for double recombinants and to entrap insertion sequences [J]. J Bacteriol, 1990. 172:3138-3145[14] Wolk C P. Physiological basis of the pattern of vegetative growth of ablue-green alga[J].Proc Natl Acad Sci USA,1967,57:1246-1251[15] Wilcox M, Mitchison G J, Smith R J. Pattern formation in the blue-green alga, Anabeana. I. Basic mechanisms [J]. J Cell Sci,1973.12:707-725[16] Wolk C P, Quine M P. Formation of one-dimensional patterns by stochastic processes and by filamentous blue-green algae [J].Develop Biol,1975. 46:370-382[17] Black T A, Cai Y, Wolk C P. Spatial expression and autoregulation of hetR, a gene involved in the control of heterocyst development in Anabaena[J].Mol Microbiol, 1993. 9:77-84

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