表达lac和glk基因的蓝藻的异养生长
HETEROTROPHIC GROWTH OF CYANOBACTERIA EXPRESSING LAC AND GLK GENES
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摘要: 将来自 Brucella abortus 的葡萄糖激酶基因 glk 和大肠杆菌乳糖操纵子基因 lacZYA 装置于蓝藻 glnA 启动子下游,以广宿主质粒导入3种蓝藻.基因的表达通过β-半乳糖苷酶和葡糖激酶活性测定得到确证.基因工程藻株均获得了利用乳糖异养生长的能力.集胞藻6803转基因株可在光照加 DCMU 的条件下利用乳糖生长,而鱼腥藻转基因株依赖乳糖可在完全黑暗条件下生长,并可在黑暗或加 DCMU 条件下诱导产生异形胞.Abstract: The glucose kinase gene glk from Brucella abortus and lacZYA genes from E. coli were positioned downstream of a cyanobacterial glnA promoter, and introduced into three cyanobacterial species in a broad-host-range plasmid. Expression of the exogenous genes was confirmed by assay of β-galactosidase and glucokinase activity. The genetically-engineered cyanobacterial strains acquired capability of heterotrophic growth on lactose. Synechocystis sp. PCC 6803(pRL2224)can grow on lactose in the light with DCMU, while Anabaena strains harboring pRL2224 are capable of heterotrophic growth in complete darkness and produce heterocysts upon induction in darkness or in the light with DCMU.
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Keywords:
- lac /
- glk /
- Cyanobacteria /
- Heterotrophic growth
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[1] Carr N G, Whitton B A. The Biology of Cyanobacteria [M]. Oxford London, Edinburgh, Boston, Melbourne: Blackwell Scientific Publications. 1982.47-85[2] Anderson S L, Mclntosh L. Light-activated heterotrophic growth of the cyanobacterium Synechocystis sp. Strain PCC 6803: a blue-light-requiring process [J]. J Bacteriol, 1991, 173: 2761-2767[3] Zhang C C, Jeanjean R, Joset F. Obligate phototrophy in cyanobacteria: more than a lack of sugar transport [J]. FEMS Microbiol Lett, 1998,161:285-292[4] Cohen M F, Wallis J G, Campbell E L, et al. Transposon mutagenesis of Nostoc sp. Strain ATCC 29133, a cyanobacterium with multiple cellular differentiation alternatives [J]. Microbiology,1994,140(Pt 12):7324-7329[5] Elhai J, Wolk C P. Conjugal transfer of DNA to cyanobacteria [J]. Methods Enzymol, 1988,167:747-754[6] Wolk C P, Cai Y, Panoff J-M. Use of transposon with luciferase as a reporter to identify environmentally responsive genes in a cyanobacterium [J]. Proc Natl Acad Sci USA, 1991. 88:5355-5359[7] Lewin B. Genes VI [M]. Oxford New York, Tokyo: Oxford University Press. 1997[8] Ramasubramanian T S, Wei T-F, Golden J W. Two Anabaena sp. Strain PCC 7120 DNA-binding factors interact with vegetative cell-and heterocyst-specific genes [J]. J. Bacteriol, 1994. 176:1214-1223[9] Essenberg R C. Cloning and characterization of the glucokinase gene of Brucella abortus 19 and identification of the three other genes [J]. J Bacteriol, 1995. 177:6297-6300[10] Khudyakov I, Wolk C P. Evidence that the hanA gene coding for HU protein is essential for heterocyst differentiation in, and cyanophage A-4(L)sensitivity of, Anabaena sp. Strain PCC 7120 [J]. J.Bacteriol, 1996.178:3572-3577[11] Black T A, Wolk C P. Analysis of a Het-mutation in Anabaena sp. Strain PCC 7120 implicates a secondary metabolite in the regulation of heterocyst spacing [J]. J Bacteriol, 1994. 176:2282-2292[12] Kroos L, Kaiser D. Construction of Tn5 lac, a transposon that fuses lacZ expression to exogenous promoters, and its introduction into Myxococcus xanthus [J]. Proc Natl Acad Sci USA, 1984.81:5816-5820[13] Miller J H. Experiments in Molecular Genetics [M]. New York: Cold Spring Harbor Laboratory Press. 1972[14] Pearce J, Carr N G. The incorporation and metabolism of glucose by Anabaena variabilis [J]. J Gen Microbiol, 1969,54:451-462[15] Williams J G. Construction of specific mutations in photosystem II photosynthetic reaction center by genetic engineering methods in Synechocystis 6803 [J]. Methods Enzymol, 1988. 167:766-778[16] Xu X, Yan G, Kong R, et al. Analysis of expression of the binary toxin genes from Bacillus sphaericus in Anabaena and the potential in mosquito control [J]. Curr Microbiol, 2000. 41:352-356
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