==================================BSR47==================================
47.  Transfection or gene transformation in protozoan.
1
UI  - 87106835
AU  - Ascenzioni F ; Lipps HJ
TI  - A linear shuttle vector for yeast and the hypotrichous ciliate
      Stylonychia.
AB  - A linear plasmid was constructed in vitro using the telomeres of the rDNA
      of Tetrahymena pyriformis. These telomeres were added to a yeast circular
      vector containing an ARS sequence from Dictyostelium, the LEU2 gene of
      yeast and the neo gene from Escherichia coli Tn5 fused with a eukaryotic
      promoter. The resulting plasmid was used to transform yeast. During the
      replication of the linear plasmid in yeast it was spontaneously modified
      at the extremity by the addition of 300 bp of yeast telomeric sequence
      for each end. Total DNA prepared from yeast transformants was used to
      transform the hypotrichous ciliate Stylonychia lemnae. The same plasmid
      isolated from Stylonychia can again be replicated in yeast.
MH  - Animal ; Ciliata/*GENETICS ; Dictyostelium/GENETICS ; Escherichia Coli/
      GENETICS ; *Genetic Vectors ; Nucleic Acid Hybridization ; Plasmids ;
      Saccharomyces Cerevisiae/*GENETICS ; Support, Non-U.S. Gov't ;
      Transformation, Genetic
SO  - Gene 1986;46(1):123-6
2
UI  - 87064635
AU  - Langford CJ ; Edwards SJ ; Smith GL ; Mitchell GF ; Moss B ; Kemp DJ ;
      Anders RF
TI  - Anchoring a secreted plasmodium antigen on the surface of recombinant
      vaccinia virus-infected cells increases its immunogenicity.
AB  - We show that the subcellular location of foreign antigens expressed in
      recombinant vaccinia viruses influences their effectiveness as
      immunogens. Live recombinant viruses induced very poor antibody responses
      to a secreted repetitive plasmodial antigen (the S-antigen) in rabbits
      and mice. The poor response accords with epidemiological data suggesting
      that S-antigens are poorly immunogenic. Appending the transmembrane
      domain of a membrane immunoglobulin (immunoglobulin G1) to its carboxy
      terminus produced a hybrid S-antigen that was no longer secreted but was
      located on the surface of virus-infected cells. This recombinant virus
      elicited high antibody titers to the S-antigen. This approach will
      facilitate the use of live virus delivery systems to immunize against a
      wide range of foreign nonsurface antigens.
MH  - Amino Acid Sequence ; Animal ; Antibody Formation ; Antigens, Protozoan/
      GENETICS/*IMMUNOLOGY ; Base Sequence ; Cell Line ; *Cell Transformation,
      Viral ; DNA, Recombinant/METABOLISM ; Enzyme-Linked Immunosorbent Assay ;
      Fluorescent Antibody Technic ; Genes, Structural ; Immunoglobulins,
      Surface/GENETICS/IMMUNOLOGY ; Mice ; Plasmodium Falciparum/GENETICS/
      *IMMUNOLOGY ; Support, Non-U.S. Gov't ; Transfection ; Vaccinia Virus/
      GENETICS/*IMMUNOLOGY
SO  - Mol Cell Biol 1986 Sep;6(9):3191-9
3
UI  - 86233431
AU  - Tondravi MM ; Yao MC
TI  - Transformation of Tetrahymena thermophila by microinjection of ribosomal
      RNA genes.
AB  - The ribosomal RNA genes (rDNA) of Tetrahymena thermophila macronucleus
      exist as free linear 21-kilobase molecules that contain replication
      origins and telomeres. A mutation in this gene confers resistance to the
      antibiotic paromomycin. We have isolated rDNA from such a mutant (strain
      p2f), microinjected it into the macronucleus of a sensitive strain, and
      obtained drug-resistant cells at a frequency of 1-3%. The transformed
      cells have a distinct and stable phenotype. The rDNA of the transformants
      contains the expected sequences of the mutant rDNA as determined by
      oligonucleotide hybridization. rDNA from a different inbred line
      (C3-368), which contains heteromorphic restriction sites, has also been
      used for injection, and the results confirm the fact that the injected
      rDNA is indeed present in the transformants. Injection of rDNA from the
      C3 strains also increases the transformation frequency 5- to 10-fold and
      leads to the total replacement of the resident rDNA of the B-inbred
      strains. This is presumably due to the replication dominance of rDNA from
      the C3 strains over that of the B strains. Using this method, we have
      also been able to transform developing cells, at similar frequencies, by
      microinjecting into the macronuclear anlagen.
MH  - Alleles ; Cell Nucleus/PHYSIOLOGY ; Conjugation, Genetic ; Cytoplasm/
      PHYSIOLOGY ; Drug Resistance, Microbial ; DNA, Ribosomal/*GENETICS ;
      Genes, Structural ; Microinjections ; Paromomycin/PHARMACOLOGY ; RNA,
      Ribosomal/*GENETICS ; Support, U.S. Gov't, P.H.S. ; Tetrahymena/*GENETICS
      ; Transformation, Genetic
SO  - Proc Natl Acad Sci USA 1986 Jun;83(12):4369-73