Biological and molecular characterization of cellular differentiation in Tetrahymena vorax: a potential biocontrol protozoan.
Tetrahymena vorax (T. vorax) is an indigenous fresh water protozoan with the natural biological potential to maintain a specific aquatic microbial flora by ingesting and eliminating specific microorganism. To investigate the molecular mechanisms controlling Tetrahymena vorax (T. vorax) cellular differentiation from a small-mouth vegetative cell to a voracious large-mouth carnivore capable of ingesting prey ciliates and bacteria from aquatic environments, we use DNA subtraction and gene discovery techniques to identify and isolate T. vorax differentiation-specific genes. The physiological necessity for one newly discovered gene, SUBII-TG, was determined in vivo using an antisense oligonucleotide directed against the 5' SUBII-TG DNA sequence. The barriers to delivering antisense oligonucleotides to the cytoplasm of T. vorax were circumvented by employing a new but simple procedure of processing the oligonucleotide with the differentiation stimulus, stomatin. In these studies, the antisense oligonucleotide down-regulated SUBII-TG mRNA expression, and blocked differentiation and ingestion of prey ciliates. The ability to down-regulate SUBII-TG expression with the antisense oligonucleotide suggests that the molecular mechanisms controlling the natural biological activities of T. vorax can be manipulated to further study its cellular differentiation and potential as a biocontrol microorganism.
Green
M M
MM
Molecular and Cellular Biology Section, Department of Biological Sciences, University of Alabama, Tuscaloosa, Alabama 35487, USA. Mgreen3@ufl.edu
LeBoeuf
R D
RD
Churchill
P F
PF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary; D018507:Gene Expression Regulation, Developmental; D009693:Nucleic Acid Hybridization / Q000379:methods; D016376:Oligonucleotides, Antisense / Q000378:metabolism; D010572:Pest Control, Biological; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013768:Tetrahymena / Q000235:genetics* / Q000502:physiology*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D016376:Oligonucleotides, Antisense; D015800:Protozoan Proteins; D012333:RNA, Messenger; C433877:SubII-TG protein, Tetrahymena vorax