Linear to radial polarization conversion in the THz domain using a passive system.

This paper addresses a passive system capable of converting a linearly polarized THz beam into a radially polarized one. This is obtained by extending to THz frequencies and waveguides an already proven concept based on mode selection in optical fibers. The approach is validated at 0.1 THz owing to the realization of a prototype involving a circular waveguide and two tapers that exhibits a radially polarized beam at its output. By a simple homothetic size reduction, the system can be easily adapted to higher THz frequencies.

Homologous recombination promoted by reverse transcriptase during copying of two distinct RNA templates.

Retroviruses are known to mutate at high rates. An important source of genetic variability is recombination taking place during reverse transcription of internal regions of the two genomic RNAs. We have designed an in vitro model system, involving genetic markers carried on two RNA templates, to allow a search for individual recombination events and to score their frequency of occurrence. We show that Moloney murine leukemia virus reverse transcriptase alone promotes homologous recombination efficiently. While RNA concentration has little effect on recombination frequency, there is a clear correlation between the amount of reverse transcriptase used in the assay and the extent of recombination observed. Under conditions mimicking the in vivo situation, a rate compatible with ex vivo estimates has been obtained.

The spread of a replication-competent MuLV retroviral vector can be efficiently blocked by deletion variants.

A retroviral vector in which the gag and pol genes have been replaced by the NLS-lacZ reporter gene was derived from a cloned AKV-like virus. A complementing cell line expressing the gag and pol retroviral genes was constructed. The retroviral vector was demonstrated to replicate in the complementing cells. Since transfection is known to generate deletion variants of the introduced plasmid, we have examined whether it can give rise to viral forms with a replicating advantage over the initial vector. After transfection in complementing cells the spread of the vector was followed by X-gal staining. The fraction of stained cells increased for the first 10 days following transfection and was then stabilized to about 20% stained cells, thus defining two cell types; one with LacZ+ phenotype and one with LacZ- phenotype. Molecular analysis showed that the latter contains a deleted form of the virus preventing cell infection by the vector presumably through a mechanism of interference involving the viral env gene. Thus, interference results in the efficient block of vector expansion.

The mammalian genome shaping activity of reverse transcriptase.

Reverse transcriptase catalyses the conversion of RNA into DNA. This operation seems to have largely contributed to the evolution of complex genomes. More than 10% of a mammalian genome is composed of sequences with reverse transcribed origin, most of which consists of repeated sequences (SINEs, LINEs). In spite of their simplicity, these sequences can play a key role in evolution by favoring illegitimate recombination. In addition to this abundant material, retrotransposed sequences include retrotransposons, retroviruses and genes depleted from intervening sequences, known as pseudogenes. Some of these sequences can be functional or involved in the regulation of neighbouring genes. These hallmarks of reverse transcription activity indicate that it has largely contributed to the fluidity of modern genomes.

Analysis of variability among endogenous ecotropic MuLV loci in laboratory mice.

We have isolated a molecular clone of an ecotropic murine leukemia virus from the ovaries of an SWR/J x RF/J hybrid female. The molecularly cloned virus, named pSR3, was demonstrated to induce virus production upon transfection into SWR/J immortalized fibroblasts and to promote germ line integration of proviruses in a fraction of the offspring germline when inoculated to neonate SWR/J females. Sequence analysis reveals that pSR3 is closely related to p623, a plasmid derived from Emv-11 (also referred to as AKV-1). Alignment of the pSR3 sequence with the partial nucleotide sequence of Emv-11 (an endogenous virus carried by BALB/c and C3H/He mice) together with p623 then allows a comparison between three viral sequences. Analysis of these data gives (a) an estimation of the natural divergence rate of MuLV genomes in the course of viral replication (1-5 x 10(-5) mutations per cycle and per nucleotide) and (b) molecular evidence for a recent origin through germ line infection of endogenous loci. From additional clues, Emv-11 appears to be the probable ancestor of at least some of these loci.

Different regulation of class I gene expression in the adult mouse and during development.

The expression of the class I genes encoding for histocompatibility Ag is complex both in adult and during development. Although ubiquitously expressed in the adult, the mRNA level of class I genes is variable from one organ to another. During development, H-2K mRNA expression has two phases: the first from blastocyst to day 11, where H-2K mRNA level is extremely low, and the second, beginning after day 11, when H-2K mRNA expression increases first dramatically (10x) and then progressively to birth. To localize the sequences responsible for the regulation of H-2K gene expression in the adult and during development, we have constructed a series of transgenic strains carrying 1) a 9-kb native H-2K gene, H-2K LF, corresponding to the entire H-2Kb gene with 2 kb of upstream sequences and 3 kb of downstream sequences, and 2) two hybrid constructs linking the same 5'-flanking region of H-2Kb gene to two reporter genes, the human growth hormone and the human c-myc proto-oncogene. Expression of the transgenes was compared with that of the endogeneous H-2K gene in adult organs and during development of the different transgenic strains. In the adult, the three constructs behave almost like the endogeneous H-2K gene, but the H-2K LF construct is the only one whose expression is independent of the integration site and related to the copy number. During development, both fusion genes are barely expressed in the embryo as well as in the extra-embryonic tissues, whereas the H-2K LF transgene expression parallels that of the endogeneous class I gene. Therefore, our results show that H-2K developmental regulatory sequences are not included in the region that controls H-2K mRNA expression in the adult, indicating that H-2K class I gene expression in adult organs and in development is regulated by different mechanisms.