The Chlorella hexose/H+ symporter is a useful selectable marker and biochemical reagent when expressed in Volvox.
The multicellular obligately photoautotrophic alga Volvox is composed of only two types of cells, somatic and reproductive. Therefore, Volvox provides the simplest model system for the study of multicellularity. Metabolic labeling experiments using radioactive precursors are crucial for the detection of stage- and cell-type-specific proteins, glycoproteins, lipids, and carbohydrates. However, wild-type Volvox lacks import systems for sugars or amino acids. To circumvent this problem, the hexose/H+ symporter (HUP1) gene from the unicellular alga Chlorella was placed under the control of the constitutive Volvox beta-tubulin promoter. The corresponding transgenic Volvox strain synthesized the sugar transporter in a functional state and was able to efficiently incorporate 14C from labeled glucose or glucosamine. Sensitivity toward the toxic glucose/mannose analogue 2-deoxy-glucose increased by orders of magnitude in transformants. Thus we report the successful transformation of Volvox with a gene of heterologous origin. The chimeric gene may be selected for in either a positive or a negative manner, because transformants exhibit both prolonged survival in the dark in the presence of glucose and greatly increased sensitivity to the toxic sugar 2-deoxyglucose. The former trait may make the gene useful as a dominant selectable marker for use in transformation studies, whereas the latter trait may make it useful in development of a gene-targeting system.
Hallmann
A
A
Lehrstuhl Biochemic I, Universität Regensburg, Germany.
Sumper
M
M
D000606:Amino Sugars; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D050260:Carbohydrate Metabolism; D002352:Carrier Proteins / Q000235:genetics*; D002708:Chlorella / Q000235:genetics*; D003001:Cloning, Molecular; D003624:Darkness; D003847:Deoxyglucose / Q000378:metabolism; D005819:Genetic Markers; D005944:Glucosamine / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins; D016133:Polymerase Chain Reaction; D012641:Selection, Genetic; D017422:Sequence Analysis, DNA; D027981:Symporters; D014170:Transformation, Genetic; D016823:Volvocida / Q000235:genetics*
D000606:Amino Sugars; D002352:Carrier Proteins; D005819:Genetic Markers; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D027981:Symporters; C089820:monosaccharide-hydrogen cotransporter; D003847:Deoxyglucose; D005947:Glucose; D005944:Glucosamine