title: Gene transfer in Leptolyngbya sp. strain BL0902, a cyanobacterium suitable for production of biomass and bioproducts.
abstract: Current cyanobacterial model organisms were not selected for their growth traits or potential for the production of renewable biomass, biofuels, or other products. The cyanobacterium strain BL0902 emerged from a search for strains with superior growth traits. Morphology and 16S rRNA sequence placed strain BL0902 in the genus Leptolyngbya. Leptolyngbya sp. strain BL0902 (hereafter Leptolyngbya BL0902) showed robust growth at temperatures from 22°C to 40°C and tolerated up to 0.5 M NaCl, 32 mM urea, high pH, and high solar irradiance. Its growth rate under outdoor conditions rivaled Arthrospira ("pirulina" strains. Leptolyngbya BL0902 accumulated higher lipid content and a higher proportion of monounsaturated fatty acids than Arthrospira strains. In addition to these desirable qualities, Leptolyngbya BL0902 is amenable to genetic engineering that is reliable, efficient, and stable. We demonstrated conjugal transfer from Escherichia coli of a plasmid based on RSF1010 and expression of spectinomycin/streptomycin resistance and yemGFP reporter transgenes. Conjugation efficiency was investigated in biparental and triparental matings with and without a "elper"plasmid that carries DNA methyltransferase genes, and with two different conjugal plasmids. We also showed that Leptolyngbya BL0902 is amenable to transposon mutagenesis with a Tn5 derivative. To facilitate genetic manipulation of Leptolyngbya BL0902, a conjugal plasmid vector was engineered to carry a trc promoter upstream of a Gateway recombination cassette. These growth properties and genetic tools position Leptolyngbya BL0902 as a model cyanobacterial production strain.
authors.affiliation: Division of Biological Sciences, University of California San Diego, La Jolla, California, United States of America.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Taton
authors.forename: Ewa
authors.initials: E
authors.lastname: Lis
authors.forename: Dawn M
authors.initials: DM
authors.lastname: Adin
authors.forename: Guogang
authors.initials: G
authors.lastname: Dong
authors.forename: Scott
authors.initials: S
authors.lastname: Cookson
authors.forename: Steve A
authors.initials: SA
authors.lastname: Kay
authors.forename: Susan S
authors.initials: SS
authors.lastname: Golden
authors.forename: James W
authors.initials: JW
authors.lastname: Golden
mesh_terms: D056804:Biofuels; D001688:Biological Products / Q000378:metabolism*; D018533:Biomass; D003227:Conjugation, Genetic; D000458:Cyanobacteria / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004526:Efficiency; D005227:Fatty Acids / Q000378:metabolism; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods; D008826:Microbial Sensitivity Tests; D008828:Microbiological Techniques; D008962:Models, Theoretical; D030781:Organisms, Genetically Modified; D010957:Plasmids; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000235:genetics
chemical_list: D056804:Biofuels; D001688:Biological Products; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D005227:Fatty Acids; D012336:RNA, Ribosomal, 16S